REVIEWS

Exporting RNA from the nucleus

to the cytoplasm
Alwin Köhler*‡ and Ed Hurt*
Abstract | The transport of RNA molecules from the nucleus to the cytoplasm is fundamental
for gene expression. The different RNA species that are produced in the nucleus are
exported through the nuclear pore complexes via mobile export receptors. Small RNAs
(such as tRNAs and microRNAs) follow relatively simple export routes by binding directly
to export receptors. Large RNAs (such as ribosomal RNAs and mRNAs) assemble into
complicated ribonucleoprotein (RNP) particles and recruit their exporters via class-specific
adaptor proteins. Export of mRNAs is unique as it is extensively coupled to transcription
(in yeast) and splicing (in metazoa). Understanding the mechanisms that connect RNP
formation with export is a major challenge in the field.

The appearance of the cell nucleus marked a central evo­ them to export RNAs. Typically, karyopherins that
lutionary transition, but the sequence of events that led import cargo are called importins and karyopherins
to this development and the immediate adaptive bene­ that export cargo are called exportins.
fits it provided to primordial eukaryotic cells remain A feature of karyopherins is their regulation by the
mysterious. Regardless of which selective pressure small GTPase Ran5. Ran exists in a GTP-bound state in
forced the nucleus–cytoplasm compartmentalization, the nucleus and a GDP-bound state in the cytoplasm.
it seems clear that by departing from the prokaryotic The RanGTP–RanGDP gradient across the nuclear
mechanism of co-transcriptional translation several membrane is generated by the action of two regu­lators,
immediate pro­blems emerged. Following the physical RanGEF (Ran-GDP-exchange factor) in the nucleus
separation of the transcription and translation pro­ and RanGAP (Ran-GTPase-activating protein) in the
cesses, a strong selective pressure must have triggered cytoplasm, and creates a driving force for directional
the co-evolution of nuclear pore complexes (NPCs) nucleocytoplasmic transport processes3. Importins bind
and nucleocytoplasmic transport machineries to allow cargo in the cytoplasm and release it after transport into
the transport of a myriad of molecules (proteins, RNAs the nucleus upon binding of RanGTP. On the other
and ribonucleoprotein (RNP) particles) between the hand, exportins bind nuclear cargo only together with
nucleus and cytoplasm. Studies over the past 2 decades RanGTP, and this ternary complex is translocated to
have revealed that nucleocytoplasmic transport occurs the cytoplasm, where it dissociates upon hydrolysis of
by various mechanisms that have in common the use RanGTP by RanGAP.
of mobile transport receptors, which bind and move Export of tRNA, microRNA (miRNA), small nuclear
diverse cargoes. These transporters can pass through the (sn)RNA and ribosomal (r)RNA follows this general
*Biochemie-Zentrum der
highly specialized channels in the nuclear membrane paradigm that involves exportins of the karyopherin
Universität Heidelberg, that are formed by NPCs1–3 (BOX 1). family and the Ran cycle6. However, general mRNA
Im Neuenheimer Feld 328, A general paradigm for nucleocytoplasmic trans­ export is mechanistically different as it uses a transport
69120 Heidelberg, Germany. port was established through the analysis of protein receptor that is unrelated to karyopherins and does not
‡
Universitätskinderklinik
import into and export from the nucleus. These studies directly depend on the RanGTP–RanGDP gradient7,8.
Heidelberg, Im Neuenheimer
Feld 151, 69120 Heidelberg, revealed that transport through NPCs requires a family Moreover, numerous additional export factors (for
Germany. of conserved nuclear transport receptors (also known as example, adaptors and release factors) cooperate with
Correspondence to E.H. karyopherins or importin‑β family members), which rec­ the mRNA export receptor6,9–11.
e‑mail: ognize a short peptide signal on a cargo protein — either In this review, we provide an overview of the major
ed.hurt@bzh.uni-heidelberg.de
doi:10.1038/nrm2255
a nuclear localization signal (NLS) or a nuclear export RNA export pathways (FIG. 1) starting with the simpler
Published online signal (NES)2–4. Moreover, karyopherins can recognize export routes of small RNAs and then discussing the
5 September 2007 nucleotide motifs in RNA cargoes, which also enables sophisticated biogenesis and export mechanisms of

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REVIEWS

Box 1 | NPC organization and nucleoporins with a role in RNA export much larger RNP particles. Minor export pathways of
individual RNAs like the signal recognition particle 7S RNA
Metazoa Yeast will not be covered. We do not discuss the nuclear export
of viral RNAs in depth (the reader is referred to Ref. 12),
mRNA mRNA mRNA but refer to them whenever they illuminate the function
snRNA of a host system that has been exploited.
mRNA TPR lp2 mRNA
1–M
rRNA Mlp Export of tRNAs
Nup60 Aminoacylated tRNAs are needed in the cytoplasm for
mRNA
NUP153 Nup1 60S, 40S ribosomal translation. About 40 different tRNAs exist
ribosomal
mRNA
subunits
in eukaryotic cells, and these tRNAs are short in length
and contain single-stranded loops and double-stranded
NUP107 Nup84 mRNA, 60S
complex complex ribosomal minihelix regions that fold into uniform cloverleaf
Nucleus FG subunits structures. tRNAs are synthesized as larger precursors
FG in the nucleus by RNA polymerase III (Pol III) (FIG. 2).
FG Nup57
Subsequent RNA-processing steps include removal of
FG Nup49 the 5′ and 3′ trailer sequences, CCA-nucleotide addi­
FG Nsp1 tion to the 3′ end to form the amino-acid acceptor stem,
FG
FG removal of introns (when present) by a specific tRNA-
FG
Cytoplasm FG splicing endonuclease and numerous base modifications
NUP107 Nup84 mRNA by tRNA-modifying enzymes13. After RNA processing
complex complex 60S, 40S and maturation, tRNAs are exported to the cytoplasm.
ribosomal Studies in several organisms established that fully pro­
mRNA RAE1 subunits
Gle2 cessed, mature tRNAs are preferentially selected by
snRNA NUP98 mRNA
Nup116
mRNA tRNA
the tRNA export machinery14,15. However, although it
rRNA NUP62 NUP88 rRNA was initially suspected that tRNA splicing occurs in the
Nsp1 Nup82
NUP214 nucleus before export, a recent study showed that the
CG1 Nup42 Nup159
mRNA mRNA yeast tRNA-splicing endonuclease is associated with
60S GLE1 Dbp5 60S, 40S the outer mitochondrial membrane16. Unexpectedly, two
Gle1 ribosomal
ribosomal DBP5 other studies reported that mature, cytoplasmic tRNAs
subunit NUP358 subunits
can be re-imported into the nucleus17,18. The implica­
mRNA mRNA tions of these findings are currently unclear, although a
function in tRNA quality control has been suggested.
mRNA mRNA
The classical tRNA export route. Export of tRNA follows
Nuclear pore complexes (NPCs) are large assemblies (~125 nm in diameter, with a mass
Nature Reviews Molecular the general paradigm of exportin-mediated protein export.
of 125 MDa in metazoa and 60 MDa in yeast) that are embedded in| the nuclearCell Biology
envelope. NPCs have an eightfold symmetrical core structure (called the spoke The class-specific tRNA export receptor exportin-t,
complex) that is sandwiched between a cytoplasmic and a nuclear ring158. The 8 spoke a member of the karyopherin superfamily, binds directly
units surround the centre of the NPC through which active transport takes place. A to tRNAs in a RanGTP-dependent manner19,20 (FIG. 2;
structure called the nuclear basket and 8 short cytoplasmic fibrils (only 4 are shown) are Supplementary information S1 (table)). The NESs in the
attached, respectively, to the nuclear and cytoplasmic ring of the NPC (see figure). tRNAs that are decoded by exportin‑t are not linear motifs,
The NPC is formed by ~30 different nuclear pore proteins (nucleoporins) that exist in but rather are coded in secondary and tertiary structural
8 or 16 (or sometimes 32) copies per NPC159,160. Nucleoporins are grouped into three elements (such as minihelices) and pro­perly processed
major classes. The first class are the FG nucleoporins, which contain Phe-Gly-rich repeat 5′ and 3′ termini, which suggests that exportin‑t can
domains that fill up the active transport channel and function directly in monitor the correct structural integrity of tRNAs before
nucleocytoplasmic transport by mediating the passage of the soluble transport
export15,21. However, exportin‑t does not discriminate
receptors161,162. The second class are nucleoporins that are devoid of FG-repeat
sequences; these are the predominant structural constituents of the NPC. The third between intron-containing and spliced tRNAs15,21. After
class are Nups, which are integral membrane proteins and are thought to anchor the transport of the tRNA–exportin‑t–RanGTP complex to
NPC in the nuclear membrane. FG nucleoporins can interact directly with the shuttling the cytoplasm, RanGAP stimulates GTP hydro­lysis on
transport receptors161. Hydrophobic patches on the surface of these transporters bind Ran, which induces release of the tRNA cargo from its
transiently to the Phe residues that are part of the FG nucleoporin network in the active receptor. Exportin‑t is the principal tRNA exporter in
transport channel161,163. vertebrate cells, but exportin-5, another member of the
Most of the nucleoporins are located symmetrically on both sides of the NPC, but a karyopherin family, is thought to be an auxiliary recep­
few nucleoporins are found asymmetrically either on the cytoplasmic or nuclear side of tor22,23. The main role of exportin‑5, however, is to export
the NPC. The asymmetrically located nucleoporins are thought to be involved in miRNAs (see below).
directional transport processes (initial receptor targeting or termination of transport) or
to fulfil compartment-specific functions at the NPC (for example, interacting with
chromatin or the transcription machinery, or as checkpoint proteins for quality-control Additional tRNA export routes. Notably, the yeast tRNA
steps during cargo export). Nucleoporins, which have been implicated in RNA export, export receptor Los1, an orthologue of exportin‑t24, is
are indicated (see main text for details). snRNA, small nuclear RNA; rRNA, ribosomal not essential for viability. This finding supported the idea
RNA. Orthologous proteins or complexes between yeast and metazoa are shown in the that Los1-independent tRNA nuclear export pathways
same colour. could exist. One of these alternative tRNA export routes

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 REVIEWS

AA
AA

Int
ro
n
5′ 3′ AAAA 3′ Cap 5′
3′
Cap Cap
tRNA miRNA snRNA mRNA rRNA

Processing, maturation and assembly with export factors

AA
AA
Mex67
Arx1
Mtr2
ALY/Yra1
CBC
Ran Ran CBC PHAX Ran Nmd3 Ran
TAP/Mex67
GTP GTP GTP GTP
CRM1 p15/ CRM1
Exp-t Exp-5 Mtr2

Nucleus

Cytoplasm
Figure 1 | The different RNA export pathways. The major RNA export routes are shown (tRNA, microRNA (miRNA),
small nuclear (sn)RNA, mRNA, ribosomal (r)RNA). In each case, the primary RNA transcript is shown,
Nature Reviewsas well as the
| Molecular Cell Biology
transport-competent RNA after it has undergone processing, maturation and assembly with export factors (export
adaptors are shown in blue, export receptors are shown in yellow). Prominent structural motifs in pre-RNAs are indicated
(single/double-stranded RNA, loops, exons and introns, 5′ cap and 3′ poly(A) tail). For the mRNA export route, the names
of both metazoan and yeast proteins are indicated, and mRNAs are shown with additional adaptor proteins and RNA-
binding factors (orange ovals). In the case of rRNA, the general exporter in eukaryotes, CRM1, and two auxiliary exporters,
Mex67–Mtr2 and Arx1, that have only been studied in yeast are depicted. CBC, cap-binding complex; Exp, exportin.

is linked to the aminoacylation machinery (FIG. 2). A to single-stranded RNA species that induce post-
study in yeast showed that the function of the Los1 tRNA transcriptional gene silencing through base-pairing
export receptor became essential when the act­ivity of an with their target mRNAs in the cytoplasm. miRNAs
aminoacyl-tRNA synthetase cofactor (Arc1) was regulate a wide range of biological processes, including
impaired25. Additional studies in Xenopus laevis oocytes developmental timing, cell differentiation, apoptosis and
and yeast demonstrated that aminoacylation of tRNAs immunity against viruses31,32. miRNA genes comprise an
can occur in the nucleus and, accordingly, aminoacylated abundant class of regulatory molecules in higher eukary­
tRNAs are exported from the nucleus to the cytoplasm26–28. otes, accounting for ~1% of the genome31, and have been
Moreover, factors of the translational machinery were estimated to target a third of all human genes33.
Signal recognition particle
shown to be required for efficient tRNA export in vivo13.
7S RNA
The signal recognition particle Taken together, these different findings point to the Biogenesis of miRNAs. The genes that encode miRNAs
is an evolutionarily conserved existence of another tRNA export pathway that overlaps can be transcribed by either Pol II or Pol III34–36 (FIG. 2).
RNA–protein complex that with the Los1-dependent route. However, no clear export The primary transcript derived from Pol II (pri-miRNA)
contains a 7S RNA species and receptor candidate has as yet been identified27,29. Recently, transiently receives a 5′ cap and a poly(A) tail similar to
targets integral membrane and
secretory proteins to the
novel auxiliary export factors that function upstream and that of mRNAs. During subsequent miRNA processing,
translocation machinery of the downstream of the tRNA-transport step through the however, the cap and poly(A) tail are removed, whereas
endoplasmic reticulum. NPC were described. One of them, Cex1, was suggested mRNAs keep these modifications. Some miRNAs can
to deliver the aminoacylated tRNAs from the nuclear also be excised co-transcriptionally from the introns
5′ cap
export receptor at the cytoplasmic side of the NPC to the of coding genes. A characteristic metazoan miRNA
A structure at the 5′ end of
eukaryotic mRNAs that ribosomal elongation factor eEF1α30. primary transcript (pri-miRNA) contains a stem with
consists of the m7GpppN cap a terminal loop and flanking segments. The stem–loop
(in which m7G represents Export of miRNAs harbours a ~22-nucleotide miRNA motif that has to be
7‑methylguanylate and ppp miRNAs, a class of non-coding RNAs that participate excised, exported and finally assembled into the RNA
represents an unusual 5′→5′
triphosphate linkage from m7G
in gene regulation, are exported by the karyopherin interference (RNAi) effector complex (FIG. 2). The stem–
to N, which is the first regular exportin‑5. miRNAs are produced as larger precursors loop is cut out (this is called cropping) in the nucleus
nucleotide of the mRNA). in the nucleus and eventually mature in the cytoplasm by a type III RNase called Drosha37, which cooperates

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a tRNA export b miRNA export

RNA Pol III miRNA in intron RNA Pol II
of gene miRNA-coding
tRNA-coding gene gene

Primary transcript Primary transcript

Intron

Exon Exon
5′ 3′ 5′ 3′ AAAA AAAA
Cap Intron Cap

5′ and 3′ processing,
base modifications, Drosha
tRNA CCA addition Splicing
Intron AAAA DGCR8
Cap Spliced mRNA

C C C
C C C Pre-miRNA
A A A
Intron-containing tRNA aa
pre-tRNA Aminoacylated
tRNA
Ran
Ran GTP
GTP
Exportin-5
Exportin-t/Los1
(exportin-5/Msn5,
other pathways?)
Nucleus Nucleus

Cytoplasm Cytoplasm

Cex1
eEF1α
Exportin-t Exportin-5

Splicing Ran Ran

GDP Dicer GDP
Aminoacylation aa
tRNA-splicing
endonuclease
Translation

Cap miRNA duplex RISC

A
Mitochondrion AAA

Figure 2 | Nuclear export of tRNA and microRNA. a | Transcription of a tRNA gene by RNA polymerase III (Pol III)
Nature Reviews | Molecular Cell Biology
generates a primary transcript, which in some cases contains an intron, with 5′ and 3′ extensions. After 5′ and 3′
processing, base modifications (red circles) and CCA-nucleotide addition at the 3′ end, the resulting tRNAs can follow
different export routes. Intron-containing and intron-free tRNAs (aminoacylated or not) are exported either by the
exportin‑t/Los1-dependent pathway, or by other less well characterized routes. After export to the cytoplasm and
dissociation of the tRNA cargo from its receptor, intron-containing tRNA is spliced by the tRNA-splicing machinery, which
is bound to the outer mitochondrial membrane in yeast. Mature tRNA is then channelled to the translational machinery by
release factors such as Cex1 and eEF1α. b | For microRNA (miRNA), only the Pol II‑dependent generation of the primary
mRNA (pri-mRNAs) is depicted. miRNAs can be encoded by an miRNA gene situated in the intron of a coding gene (left
branch) or from a separate miRNA gene (right branch). In both cases, the primary transcript is cleaved by the Drosha–
DGCR8 complex to generate a pre-miRNA with a stem–loop structure. Moreover, a spliced mRNA can be generated
concomitantly with the excision of an intronic miRNA. The pre-miRNA with its typical ~2-nucleotide overhang at its 3′
end is specifically recognized by exportin‑5 and is transported to the cytoplasm, where it dissociates from its receptor
after RanGTP hydrolysis. Following release from exportin‑5, Dicer further cleaves the pre-miRNA to finally generate a
single-stranded miRNA species, which assembles into the RNA-induced silencing complex (RISC). Highlighted in blue is
the RNA region that corresponds to the mature effector miRNA.

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with the RNA-binding protein DGCR838. DGCR8 is Spliceosomal snRNP biogenesis. The spliceosomal snRNAs
thought to function as a molecular ruler that measures participate in the removal of introns from pre-mRNAs.
the distance from the base of the stem and thus positions With the exception of U6 snRNA, which is produced by
Drosha precisely for cleavage38. Pol III and does not exit from the nucleus, all the other
The released ~65-nucleotide stem–loop intermedi­ spliceosomal snRNAs (U1, U2, U4 and U5) are synthe­
ate (now called pre-miRNA) is subsequently exported sized as precursors (pre-snRNAs) by Pol II and acquire
to the cytoplasm in a RanGTP-dependent manner by a 5′ cap that constitutes the signal for nuclear export
exportin‑5, a member of the karyopherin family39–41 (FIG. 1). However, pre-snRNAs do not become polyadenyl­
(FIG. 2). After release in the cytoplasm upon GTP hydrol­ ated, although they exhibit specific 3′-end processing
ysis on Ran, the pre-miRNA hairpin is further cleaved that requires factors related, but not identical, to the
by Dicer, another type III RNase that produces a ~22- 3′-end processing machinery that acts on pre-mRNAs52.
nucleotide miRNA duplex42 that contains imperfect base Why snRNA is exported from the nucleus before being
pairings. These mismatches cause one duplex strand imported again to function in splicing is still unknown,
to be less stably base paired at its 5′ end, which leads to but it was proposed that the cytoplasmic phase of snRNA
its degradation, while the other strand is incorporated biogenesis might provide a proofreading step to prevent
into an effector complex known as the RNA-induced nuclear accumulation of non-functional snRNAs53.
silencing complex (RISC)42–44. RISC finally binds to its
target mRNA through base pairing of the miRNA to Phosphorylation-dependent snRNA export. The export
the 3′ untranslated region (3′ UTR), thereby inducing receptor for snRNA is CRM1 (also known as exportin‑1),
mRNA cleavage, translational inhibition by sequester­ the general protein exporter that recognizes the Leu-
ing the translational apparatus in P bodies or cleavage- rich-type NES on proteins54,55. Consistently, CRM1 does
independent mRNA degradation45,46. not directly interact with the snRNA cargo, but requires
the cap-binding complex (CBC) and a NES-containing
Coupling of miRNA processing with export. Biogenesis and adaptor protein called PHAX to be targeted to the
nuclear export of miRNAs are coupled at several levels32. 5′ cap of the snRNA56–58. Phosphorylation of PHAX in
The key enzyme involved in this coupling is Drosha, the nucleus is required for recruitment of CRM1 and
which generates a double-stranded RNA minihelix with a RanGTP to the CBC-bound snRNA complex (FIG. 1).
P bodies ~2-nucleotide 3′ overhang, the unique structure of which After export to the cytoplasm, GTP hydrolysis of Ran and
(Processing bodies). Discrete is recognized both by exportin‑5 and the downstream- dephosphorylation of PHAX are necessary to efficiently
cytoplasmic foci that are sites acting processing enzyme Dicer. Thus, a strict linkage of dissociate the export complex and release the snRNA58.
of degradation and surveillance all processing and export steps ensures the high specificity Once in the cytoplasm, the survival of motor neurons (SMN)
of mRNAs and RNA-mediated
gene silencing.
of miRNA production and function (FIG. 2). complex facilitates the assembly of the exported snRNA
Notably, many human miRNA genes are embedded with a heteroheptameric ring of Sm proteins, which bind
Short hairpin RNA in the intronic regions of coding genes and thus use an to a conserved Sm-binding site that is present on each
Artificially generated, usually un­usual biogenesis pathway, as their production is coupled snRNA53. Binding of Sm proteins induces trimethylation
vector-encoded, RNA that
to mRNA splicing 47–49 (FIG. 2) . It seems that Drosha of the cap and exonucleolytic removal of the 3′ trailer
resembles pre-miRNA in
structure and is used to releases the pre-miRNA from the intron shortly before sequences. The trimethylated cap and the associated Sm
experimentally induce RNA splicing, allowing the generation of both RNA species at proteins provide a composite nuclear targeting signal for
interference. the same time49. subsequent nuclear import of the mature snRNPs59. After
Nuclear export of miRNA has important implications for re-import into the nucleus, snRNPs together with numer­
Small interfering RNA
(siRNA). Short double-stranded
applied aspects of RNAi technology50, as short hairpin RNAs ous other splicing factors assemble into the functional
RNA fragment of ~22 use the same export pathway to produce small interfering spliceosome53.
nucleotides that is derived (si)RNAs51. The expression of exportin‑5 is low in most cell
from longer double-stranded types, which suggests that the miRNA export receptor Export of mRNAs
short hairpin RNA. siRNAs
could limit the desired expression of siRNAs when used RNA classes with highly related structures (tRNAs and
guide silencing complexes to
their targets by base pairing as a therapeutic tool in molecular medicine51. miRNAs) exhibit common identity elements that can
with specific mRNA sequences. be decoded by class-specific export receptors. mRNAs,
Export of snRNAs however, differ substantially in length, sequence and
Survival of motor neurons The minimalist nuclear export device for RNA requires structure, and so use other strategies to find their class-
(SMN) complex
A large multiprotein complex
‘naked’ RNA as cargo, an exportin and RanGTP. specific export receptors. mRNAs are channelled into
that brings together the Sm Although this model is sufficient to explain export of the specific export pathway coordinately with their
proteins and small nuclear tRNAs and miRNAs, it does not account for the nuclear processing and assembly into messenger (m)RNPs.
RNAs, thereby facilitating small export of other cellular RNAs, which require more Typically, eukaryotic mRNAs are synthesized by Pol II
nuclear ribonucleoprotein
elaborate transport mechanisms. In terms of complexity, as precursors (pre-mRNAs) that become capped at the
assembly.
spliceosomal snRNAs lie between tRNAs/miRNAs and 5′ end, spliced, cleaved at the 3′ end and polyadenylated
Sm proteins mRNAs/rRNAs. Their export requires adaptor proteins (FIG. 1). During the successive steps of mRNP formation,
A set of seven proteins that are that recruit the export receptor (a mechanism that is many RNA-binding and -modifying proteins (for exam­
arranged as a ring structure on extensively used in the case of mRNA and rRNA export). ple, capping, splicing and processing factors) are recruited
a specific small nuclear RNA-
binding site to become part of
However, assembly into an RNP, another hallmark of to the transcripts. Genome-wide analyses revealed a pref­
spliceosomal small nuclear mRNA and rRNA transport, is not needed for the nuclear erential association of certain RNA-binding proteins with
ribonucleoproteins. export of snRNAs. distinct functional classes of mRNAs, which suggests

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that biogenesis, export and translation of mRNA subpop­ Finally, it is well established that CRM1 acts in the
ulations may be coordinated differently 60,61. Among the nuclear export of a number of unspliced and partially
factors bound to the pre-mRNAs are also export adap­ spliced viral mRNAs. These viral mRNAs can bind adap­
tors that serve to establish a physical bridge between the tor proteins that contain NESs (for example, HIV Rev,
mRNA molecule and its export receptor. Studies carried adenovirus E1b 55 kDa), thereby targeting the transport
out in a number of different model organisms revealed receptor CRM1 (ref. 12). In fact, it was the identification
that the mRNA export pathway is conserved from yeast of the NES in HIV Rev that led to the discovery of the
to humans, although every model organism has its own CRM1-dependent nuclear protein export pathway54.
peculiarities7,9,12,62.
Practically every step of mRNA biogenesis involves The Yra1/ALY/REF adaptor. The mRNA export recep­
rigorous quality control to detect possible errors in trans­ tor is targeted to different transcripts by export adaptors
cription, mRNA processing or export63. Nuclear mRNA that are typically mRNA-binding proteins. So far, only
surveillance has been studied mostly in Saccharomyces a few of these adaptor proteins have been identified,
cerevisiae. In analogy to the cytoplasmic machineries, but it is likely that more will be uncovered among the
nuclear pre-mRNAs can be degraded from either the 5′ or huge number of predicted RNA-binding proteins in
3′ end. The actual pathway seems to depend on intrinsic eukaryotic cells.
features of the substrate (for example, the stage of bio­ The Yra1 or ALY/REF adaptor (Yra1 in yeast and
genesis, intron-containing or not, and so on). mRNA ALY or REF in metazoa) can associate directly with the
decay in the 3′→5′ direction (for example, mRNAs with general mRNA exporter7,62 (FIG. 3). Moreover, Yra1 and
defective poly(A) tails) is mediated by the exosome, a ALY/REF physically interact with another conserved
complex of exonucleases that is functionally connected to export factor — Sub2 in yeast and UAP56 in metazoa77,78.
various activating cofactors63–66. Apart from pre-mRNA Sub2 and UAP56 are RNA helicases that can associate
turnover, the exosome is also responsible for degrading with several complexes involved in mRNP biogenesis
aberrant tRNAs, snRNAs, small nucleolar RNAs and (for example, the TREX (transcription-coupled export)
rRNAs67. Another quality checkpoint is provided by the complex and the spliceosome; see below). Thus, Yra1
Mlp1–Mlp2 system, which consists of two filamentous and ALY/REF form a bridge between an upstream-acting
proteins that are attached to the nuclear face of NPCs and RNA-binding protein and a downstream-acting mRNA
prevent the exit of unspliced transcripts63,68–70. export receptor. Consistent with this model, Yra1 is
co-transcriptionally recruited to nascent transcripts and
A general and a specific mRNA receptor. The yeast is therefore present at different steps during pre-mRNA
Mex67–Mtr2 complex and the homologous metazoan formation and processing79,80.
TAP–p15 complex (also known as NXF1–NXT1) func­ In addition to its role in export of cellular mRNAs,
tion as general mRNA export receptors to transport ALY/REF can function as an adaptor for the export of
mRNPs through the NPCs71,72. Although the mRNA viral intronless mRNAs. For example, the herpes sim­
exporter can bind directly to RNA, it operates together plex virus protein ICP27 targets ALY/REF to access the
with adaptor RNA-binding proteins (FIG. 3). The con­ TAP–p15-mediated export pathway of the host cell81.
served mRNA exporter is structurally unrelated to the
karyopherins and is RanGTP independent; therefore, SR proteins can function as adaptors. The SR (Ser/Arg-rich)
directionality of transport would have to be established by proteins have essential roles in splicing but also function
other mechanisms. Nevertheless, like karyopherins, the as adaptors and regulators of multiple steps of mRNA
mRNA exporter can physically interact with the Phe-Gly- metabolism including mRNA export, stability and
rich repeats of FG nucleoporins, which allows it to over­ translation11. SR proteins are abundant, evolutionarily
come the permeability barrier of the NPC that is formed conserved phosphoproteins. After splicing, several SR
by the FG-nucleoporin meshwork7,10,62 (BOX 1). Besides proteins remain bound to the spliced transcript and are
its physiological role in cellular mRNA export, human exported to the cytoplasm, where they dissociate from
TAP transports a set of viral pre-mRNAs to the cyto­ the transcript and are re-imported82. Shuttling SR pro­
plasm by binding directly to specific viral RNA elements teins (for example, SRP20 or 9G8) can recruit the general
called constitutive transport elements (CTEs)72. mRNA export receptor TAP–p15. TAP–p15 can bind
The general RanGTP-dependent protein export directly to the SR motifs, which are sites of reversible
AU-rich element
A motif that is located in the
receptor CRM1 does not have a major role in mRNA phosphorylation83,84. SR proteins are recruited in a hyper­
3′-untranslated region of some export73,74. However, CRM1 can be involved in the nuclear phosphorylated form to the splicing machinery, but after
mRNAs and that can induce export of a subset of transcripts, such as mRNAs of sev­ splicing become hypophosphoryl­ated, which favours the
rapid mRNA decay. eral protooncogenes and cytokines, that contain AU‑rich binding of TAP–p15 (ref. 83). Thus, the phosphorylation
elements (AREs) in their 3′ UTRs75. These AREs can target status of SR proteins could act as a switch to signal the
SR (Ser/Arg-rich) proteins
An abundant class of proteins NES-containing adaptor proteins and hence become con­ export competence of the spliced mRNP.
that are involved in various nected to the CRM1-dependent export pathway. In yeast, A cycle of phosphorylation and dephosphorylation
aspects of mRNA metabolism. Crm1 was recently reported to be required for export of of an SR protein (Npl3) has also been implicated in a
They contain one or two RNA- the unspliced YRA1 pre-mRNA (Yra1 is itself an mRNA termination step during mRNA export in yeast85. Npl3 is
recognition motifs and an
Arg/Ser-rich domain that can
export adaptor; see below)76. However, the mechanistic first phosphorylated by the Sky1 kinase in the cytoplasm,
be phosphorylated at multiple details of how Crm1 in conjunction with Mex67–Mtr2 can which stimulates nuclear import of Npl3 (ref. 86). In the
positions. export an unspliced pre-mRNA are currently not clear. nucleus, Npl3 can associate with the nascent transcript in

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a Yeast b Metazoa Pol II

Pol II
Transcription Transcription

THO
TREX Sub2
Yra1
Cap E I E I E I E
Splicing CB
Cap C

AAAA
TREX THO EJC
UAP56
Recruitment CB
C ALY
and export THO Recruitment
Sub2 THO

AAAA
and export
UAP56
Mex67 Npl3
Mtr2 AAAA
TAP
CB Yra1 EJC
p15 TAP
C Mex67 p15
Mtr2 CB
C ALY SR

AAAA
AAAA

AAAA
Nucleus Nucleus

Npl3 Cytoplasm SR Cytoplasm

Gle1 Gle1
Translocation Yra1 Dbp5
Dbp5 InsP6
and termination Mex67 TAP
p15
ALY
Mtr2
CBC
CBC TAP
Release and Release and p15
Mex67
re-import Yra1 re-import
Mtr2
ALY
Translation Translation
initiation eIF4A initiation eIF4A
Npl3 SR
AAAA AAAA

Figure 3 | Transcription-coupled or splicing-coupled mRNA export. a | In yeast, the nascent transcript generated
by RNA polymerase II (Pol II) is co-transcriptionally folded and assembled into a pre-messenger ribonucleoprotein
(pre-mRNP) by the THO/TREX complex (Yra1 and Sub2 are subunits of TREX, and THONature Reviews | Molecular
is a subcomplex) Cell Biology
that accompanies
the elongating Pol II. After co-transcriptional association with additional pre-mRNA factors (for example, the cap-binding
complex (CBC) and RNA-binding proteins, shown in orange), the Mex67–Mtr2 mRNA export receptor is recruited to the
mRNP via adaptor proteins such as Yra1. When emerging at the cytoplasmic side of the nuclear pore complex (NPC),
the mRNP encounters a remodelling machinery on the NPC fibrils that consists of the ATP-dependent RNA helicase
Dbp5 and its activators Gle1 and the signalling molecule inositol hexakisphosphate (InsP6). After dissociation of Mex67–
Mtr2 and other export factors from the RNA cargo, the mRNA may still contain a few shuttling RNA-binding proteins,
such as Npl3, that influence translation. mRNA is recruited to the translation-initiation machinery via its 5′ cap that is
recognized by the initiation factor eIF4A and by additional initiation factors. Transport factors are re-imported, which in
some cases requires phosphorylation. b | For metazoa, several mRNA export models have been described. Depicted are
the splicing- and cap-dependent modes of human TREX recruitment to the mRNP. The downstream events in mRNA
export, including the recruitment of the TAP–p15 mRNA export receptor, are similar for metazoa and yeast. Export
receptors are indicated in yellow and adaptors in blue. For simplicity, the exon-junction complex (EJC) on the
translocating and cytoplasmic mRNP is not shown. E, exon; I, intron; TREX, transcription-coupled export complex.

a transcription-dependent manner87. Subsequently, Npl3 TREX in transcription-coupled export. The conserved

becomes dephosphorylated by the nuclear phosphatase TREX complex integrates steps in mRNA biogenesis with
Glc7, allowing interaction with the Mex67–Mtr2 export nuclear export. TREX is found in yeast and higher eukary­
receptor. After transport of the mRNP to the cytoplasm, otes including Drosophila melanogaster and humans88–90.
Npl3 is rephosphorylated by Sky1, which destabilizes Four of the yeast TREX subunits (Tho2, Hpr1, Mft1 and
the interaction of Npl3 with the mRNA and Mex67– Thp2) form a robust subcomplex (THO)91 that functions
Mtr2 (ref. 86). Thus, successive phosphorylation and in various aspects of co-transcriptional mRNP formation
dephosphorylation of Npl3 could be a means to impose and transcription-dependent recombination92. Two of the
directionality on the mRNA export pathway85. TREX subunits, Yra1 and Sub2, are export factors that

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REVIEWS

generate a physical link with the mRNA export receptor The TREX‑2 mRNA export complex. Sac3 was identified
(FIG. 3). In yeast, TREX is continuously loaded onto emerg­ as an additional mRNA export adaptor in yeast owing
ing transcripts during transcription elongation, which to its genetic interaction with the TREX complex and
facilitates folding of nascent transcripts into mRNPs and its ability to physically recruit the Mex67–Mtr2 export
helps to recruit additional RNA-binding proteins78,88,93–95. receptor104,105. Sac3, together with Thp1, Sus1 and Cdc31,
When yeast TREX follows the elongating RNA constitute a complex (previously called the Sac3–Thp1–
polymerase along the activated gene, members of the Sus1–Cdc31 complex)104,106–108 that we henceforth call
THO subcomplex (for example, Hpr1) bind to chromatin, TREX‑2 (FIG. 4). TREX‑2 is tethered to the inner side of
whereas Sub2 and Yra1 associate with nascent mRNA93 the NPC via the nucleoporins Nup1 and Nup60 (ref. 104).
(FIG. 3). RNAi studies in D. melanogaster and transcrip­ Interestingly, one TREX‑2 component, the small protein
tion profiling have led to the suggestion that the majority Sus1, also interacts with SAGA, a large transcription-
of mRNAs are transcribed and exported independently of initiation complex that catalyses histone acetylation
TREX89. However, TREX mutants show nuclear accumu­ and deubiquitylation. In SAGA, Sus1 is part of a hetero­
lation of bulk mRNA and are synthetically lethal when trimeric deubiquitylation module together with Sgf11 and
combined with mutants of the mRNA export machinery, the protease Ubp8 (refs 109,110). TREX‑2 has therefore
which suggests a broad role for TREX in transcription- been proposed to functionally couple SAGA-dependent
coupled mRNA export. In the absence of a functional gene expression to mRNA export at the inner side of the
THO subcomplex, the formation of an RNA–DNA hybrid NPC106. This model is supported by recent experiments
between a nascent unfolded transcript and the DNA tem­ that demonstrated a requirement for both SAGA and
plate was observed, and this resulted in the inhibition of TREX‑2 in the dynamic repositioning of gene loci from
transcription and the generation of malformed transcripts the nuclear interior to the nuclear periphery111.
that could not be exported94. However, by experimentally
slowing down transcription, these defects can be partly Gene gating and mRNA export. More than 20 years
compensated, which suggests that only a limited time win­ ago, Günter Blobel proposed in his provocative gene-
dow exists for co-transcriptional loading of RNA-binding gating hypothesis that every gene in the nucleus is
proteins and export factors onto the nascent transcript96. physically connected (or gated) to a particular NPC
Despite its conservation, the TREX complex is asso­ in the nuclear membrane112. Several recent studies in
ciated with the transcription apparatus in yeast and yeast, D. melanogaster and mice indicated that gene
the splicing machinery in humans, possibly reflecting the gating indeed exists, although not in the strict sense as
higher prevalence of spliced mRNA in mammals than originally proposed113. In yeast and higher eukaryotes,
in yeast9. Initially, studies carried out in the Reed labora­ the nuclear periphery was classically considered as
tory suggested a close link between splicing and mRNA a zone of transcriptional repression owing to the pre­
export in higher eukaryotes with a key role for ALY/REF sence of silencing factors113. However, a number of
and UAP56 in this coupling97,98. Subsequent work from genes are dynamically targeted to the nuclear enve­
several laboratories reported that ALY/REF, UAP56 and lope upon activation, and this positioning appears to
TAP–p15 can associate with the exon-junction complex facilitate transcription and subsequent nuclear mRNA
(EJC), which is deposited as a consequence of splicing export111,114–119.
~20–24 nucleotides upstream of every exon–exon junc­ The precise molecular basis for the initial targeting
tion in the spliced mRNA99,100. The EJC operates in the and subsequent tethering of genes to the nuclear periph­
nonsense-mediated decay (NMD) pathway. Additionally, ery is currently unclear, but can apparently involve multi­
due to its association with export factors, the EJC was sug­ ple factors (FIG. 4). In support of the idea that gene gating
gested to recruit the mRNA export machinery and link and mRNA export are functionally coupled, the SAGA
splicing with export in metazoa. However, a recent study transcription-initiation complex was found to generate
showed that the human TREX complex is recruited in a a physical contact with the NPC-associated TREX‑2
Exon-junction complex
splicing- and cap-dependent manner only to the 5′ end mRNA export complex106,111,120. Additional gene–NPC
(EJC). A complex of proteins
that is deposited onto mRNA of the mRNA, and this recruitment requires the cap- interactions are mediated between Mex67 and the
during pre-mRNA splicing binding subunit CBP80, which interacts directly with the Mlp proteins, between the histone variant Htz1 and
(~20–24 nucleotides ALY/REF subunit of human TREX101 (FIG. 3). This obser­ the nucleo­porin Nup2 and through the Nup84
upstream of exon–exon vation could explain why mRNA transcripts are exported complex119,121,122. These factors might either operate
junctions). The EJC remains
bound to the mRNA during
in a 5′→3′ direction102. Notably, a function for the 5′ cap synergistically or in a gene-specific manner.
nuclear export and influences and the CBC in metazoan mRNA export was already Two general mechanisms of gene gating have
surveillance, translation and proposed more than 15 years ago56. emerged: either genes can interact directly with the NPC
localization of mature mRNAs In addition, the metazoan mRNA export machinery via adaptor proteins independently of mRNA produc­
in the cytoplasm.
can be loaded onto nascent transcripts by a splicing- tion (including post-transcriptional tethering)119,122–124,
Nonsense-mediated mRNA independent mechanism similar to the one in yeast. In or transcript formation is necessary for gene gating to
decay human cells, the transcription-elongation factor SPT6 occur111,115–118,125. Taken together, gene gating could be
(NMD). A process by which a recruits IWS1 to the nascent mRNA, thereby allowing a means to create a favourable environment for the
cell destroys mRNAs for which IWS1 to act as a bridging protein for ALY/REF103. Thus, recruitment of mRNA export factors and mRNA qual­
translation has been
prematurely terminated owing
eukaryotic cells can exploit different mechanisms to load ity surveillance by the Mlp1–Mlp2 network, which in
to the presence of a nonsense mRNA-processing and export factors onto newly forming turn could increase the efficiency of mRNP entry into
codon in the coding region. mRNPs. the transport channel of the NPC106,117,120,126.

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© 2007 Nature Publishing Group
 REVIEWS

New evidence provides an explanation for how Dbp5

could dismantle the mRNP and release Mex67–Mtr2 only
in the cytoplasmic compartment132–134. According to these
studies, Dbp5 exhibits a very low ATP-dependent RNA-
Pol II helicase activity, which can be stimulated by Gle1, an
essential mRNA export factor that is also asymmetrically
located at the cytoplasmic nuclear pore filaments133,134
Tho2
SAGA Htz1-containing (FIG. 3). Maximal stimulation of the ATPase activity of
Htz1- nucleosome Mft1 Thp2 Dbp5 requires the signalling molecule inositol hexakisphos‑
containing Ubp8
Sus1 Hpr1 THO/TREX phate (InsP6), which is thought to regulate the interaction
nucleosome Sub2
Sgf11 between Gle1 and Dbp5. Previously, it was also shown
Yra1
that the signalling pathway leading to the production of
Mtr2 cytoplasmic InsP6 is necessary for Gle1-dependent mRNA
Mex67 Cap export in yeast135. Taken together, these data have rein­
Thp1
TREX-2 Sac3 forced a model of local mRNP remodelling by an RNA
Sus1 Cdc31 helicase that becomes activated at the cytoplasmic pore
2
–Mlp filaments. This local activation could generate directional­
Nup1 Mlp1
Nup2 ity either by preventing backsliding of the mRNP (like a
Nup60
Nucleus molecular ratchet) from the cytoplasm to the nucleus or
by exerting a pulling force on the protruding 5′ end of the
mRNA while in transit10,136.
Cytoplasm
Export of rRNA
Ribosomes are the protein-synthesizing machines of
Figure 4 | Gene gating and mRNA export. Model of transcription-coupled mRNA export the cell. They predominantly translate mRNAs in the
Nature Reviews | Molecular Cell Biology
in yeast, which involves gating of the activated gene to the nuclear periphery via cytoplasm, but mitochondria and chloroplasts have their
interaction with the nuclear pore complex (NPC). During transcription initiation, SAGA is own set of ribosomes. Ribosomes are composed of a large
recruited as part of a large transcription pre-initiation complex to the promoter of a gene, (60S) and a small (40S) subunit, which together contain
which at this point is located in the interior of the nucleus. Subsequently, the activated 4 rRNA species (28S/25S rRNA, 5.8S rRNA, 5S rRNA
gene becomes tethered to the nuclear periphery via an interaction between SAGA and and 18S rRNA) and more than 70 ribosomal proteins.
the nuclear pore-bound TREX‑2 complex (which consists of the subunits Sac3, Thp1,
The ribosomal subunits are assembled in the nucleolus
Sus1 and Cdc31). Transcription of the tethered gene generates a nascent transcript that
and are transported to the cytoplasm by multiple export
becomes assembled, with the aid of the THO/TREX complex, into an export-competent
messenger ribonucleoprotein (mRNP). Thus, the mRNP is brought into the vicinity of the receptors.
NPC-associated Mex67–Mtr2 mRNA export machinery. Mex67 can also directly contact
the promoter region independently of the mRNA (not shown). Sus1 is a functional Ribosome biogenesis. Ribosome biogenesis in eukaryotic
component of both TREX‑2 and SAGA. Within SAGA, Sus1 is part of a heterotrimeric cells is a highly regulated multistep process. It requires the
subcomplex together with the histone H2B deubiquitylating enzyme Ubp8 and Sgf11, transcription of precursor rRNAs (pre-rRNAs) from the
a protein of unknown function. The nucleoporin Nup1 is part of the TREX‑2 docking site ribosomal genes in the nucleolus as well as the synthesis of
at the nuclear basket. Nup2 was shown to associate with active promoters and might ribosomal proteins in the cytoplasm and the subsequent
tether genes through the histone variant Htz1 in yeast. The Mlp1 and Mlp2 proteins import of these ribosomal proteins into the nucleolus. In
contribute to RNA surveillance by retention of unspliced mRNA. Dotted lines represent
the nucleolus, the ribosomal proteins are then assembled
predicted interactions between the complexes. TREX, transcription-coupled export
with the nascent pre-rRNA. Ribosome biogenesis and
complex. Proteins in the same complex have the same colour.
export has mainly been analysed in S. cerevisiae. These
studies revealed that more than 150 non-ribosomal
factors associate transiently with the evolving pre-ribos­
Directionality and termination of mRNA export. omal particles on their way from the nucleolus through
Unidirectional movement of mRNPs from the the nucleoplasm into the cytoplasm137–139. Notably, the
nucleus into the cytoplasm requires a termination nuclear export machinery is not recruited at early stages
step to release the mRNA from its receptor. In prin­ of pre-ribosome assembly (that is, during pre-rRNA
ciple, remodelling of the mRNP upon its arrival in synthesis, processing and modification), but only at a
the cytoplasm could be a driving force for vectorial late stage, in the nucleoplasm, when the pre-ribosomal
translocation. ATP-dependent RNA helicases such particles have already undergone a complicated cascade
as Dbp5, which are involved in mRNA export, could of processing, maturation and quality control.
trigger an irreversible ATP-driven mRNP rearrange­
Inositol hexakisphosphate
(InsP6). One of many small ment at the cytoplasmic side of the NPC127,128. Dbp5 Export of the ribosomal subunits. During the early stage
messenger phosphoinositides is advantageously located to perform such a role as of ribosome biogenesis, pre‑60S and pre‑40S particles
that is found in cells. InsP6 is it is associated with the nucleo­porin Nup159, which are generated in the nucleolus, and these subsequently
synthesized by IPK1 from is asymmetrically positioned at the cytoplasmic side follow separate export routes. Currently, the mechanism
inositol 1,4,5-trisphosphate
(InsP3), a precursor that also
of the NPC129,130. However, Dbp5 shuttles between the of 40S subunit export is poorly understood. So far, it is
regulates the release of nucleus and the cytoplasm and is already recruited to only known that the Ran cycle and Crm1 play a role140.
intracellular calcium. nascent mRNPs during transcription128,129,131. Several pre‑40S assembly factors and ribosomal small

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© 2007 Nature Publishing Group
REVIEWS

Mtr2 Ran
Mtr2 GTP Mtr2 Nucleus
Mex67
Mex67 Mex67
pre-60S pre-60S pre-60S

Nmd3 Arx1 Nmd3 Arx1

Ran
Nmd3 Crm1 GTP
Mtr2 FG
Arx1 FG
Crm1 Mex67
Nucleus pre-60S FG
FG
Nmd3
Ran Arx1
Cytoplasm Crm1 GTP

Lsg1
GTP
Rpl10 Mtr2
Mex67
pre-60S pre-60S pre-60S

Rpl10 Arx1 Nmd3 Arx1 Nmd3 Arx1

Ran
Crm1 GTP

Mex67
Rei1 Lsg1 Ran Cytoplasm
Nmd3 Crm1 GDP Mtr2
GDP
Arx1 Figure 5 | Nuclear export of ribosomal subunits. The export of the pre‑60S ribosomal subunit in yeast is shown. After a
complicated series of assembly steps in the nucleolus, the pre‑60S particle reaches the nucleoplasm
Nature Reviews | and recruits
Molecular several
Cell Biology
different classes of export receptors. The Crm1 export receptor binds in the presence of RanGTP to the nuclear export
signal (NES)-containing adaptor Nmd3. The other export receptors Mex67–Mtr2 and Arx1 can bind directly to the pre‑60S
subunit. After translocation through the nuclear pore complex (NPC) channel, the export factors are removed from the
pre‑60S particle in the cytoplasm. Crm1 dissociates from Nmd3 upon RanGTP hydrolysis. Nmd3 is released by the Lsg1
60S GTPase and is replaced by the ribosomal protein Rpl10. Arx1 might be dissociated by the release factor Rei1. The inset
depicts a magnified version of the NPC channel and shows all export receptors that interact with both the pre‑60S particle
Rpl10 and FG nucleoporins. Export receptors are yellow, adaptors are blue and dissociation factors are pink.

subunit proteins have been implicated in 40S export, Arx1 is another auxiliary shuttling export factor that
but it is not clear whether they are bona fide export is recruited to the late pre‑60S particle concomitantly
adaptors141–143. with Nmd3 and Mex67–Mtr2 (refs 150,154–156) (FIG. 5).
The mechanism of nuclear exit of 60S subunits has After nuclear export, Arx1 and its interacting partner
been elucidated in yeast and metazoa. These studies Alb1 are released from the 60S subunit by Rei1, a cyto­
revealed that 60S subunit export is conserved and plasmic factor that operates at a terminal step during
depends on the Ran system and the Crm1 export recep­ pre‑60S biogenesis154,155. A recent study demonstrated
tor144–149 (FIG. 5). Nmd3, a conserved NES-containing that Arx1 has properties of shuttling transport recep­
protein that is recruited to a late export-competent tors, which can directly interact with FG nucleoporins157.
pre‑60S particle in the nucleoplasm, serves as the adaptor However, Arx1 is not related to typical nucleocytoplas­
protein146–151. Following export to the cytoplasm, Crm1 mic transport receptors, but instead is homologous to
is dissociated from the Nmd3 adaptor by RanGTP hydro­ Met aminopeptidases (MetAPs). It is speculated that
lysis. Subsequently, Nmd3 is released from the 60S export Arx1, which does not have a MetAP activity, never­theless
cargo by a cytoplasmic GTPase (Lsg1), an event that is uses its MetAP fold to bind to the Phe-Gly repeats of FG
coupled to the loading of the ribosomal protein Rpl10 nucleoporins157.
onto the 60S subunit151,152. Thus, pre‑60S subunits, in contrast to other RNA
Genetic studies in yeast recently indicated that classes, can recruit several different export receptors to
Crm1 is not the sole export receptor that escorts the make their export more efficient. Notably, 60S subunits
pre‑60S particle through the NPC. Surprisingly, one of belong to the largest RNA-containing particles that have
the additional export receptors for the pre‑60S particle to pass the transport channels of the NPCs. The overall
is Mex67–Mtr2, which uses a distinct interaction sur­ size of a 60S subunit is in the range of the functional
face to bind to the pre‑60S subunit153 (FIG. 5). Thus, the diameter of the NPC, which is ~26 nm2. It is conceiv­
Mex67–Mtr2 heterodimer provides a versatile molecu­ able that the bulky pre‑60S particle needs to mobilize
lar surface to bind to cargoes as diverse as mRNPs and several export receptors for an efficient passage through
pre-ribosomal particles. the NPC (FIG. 5).

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© 2007 Nature Publishing Group
 REVIEWS

Concluding remarks yet developed the sophisticated in vitro export assays

Numerous studies from the past 20 years have shed light that are needed to recapitulate upstream events that
on how the major RNA species are exported from the occur during RNA generation, processing and RNP
nucleus into the cytoplasm. Today, we have detailed assembly.
information about the factors that are involved in Deciphering the functional connectivity of the differ­
RNA export, and we are steadily gaining mechanistic ent steps in RNA biogenesis in relation to nuclear export
insight into RNA-export processes. However, we do not is therefore still a major challenge. Hypothetically, the
have a profound structural knowledge of RNA export. combinatorial use of nuclear RNA-binding proteins
Moreover, the lack of a comprehensive mechanistic might significantly expand the regulatory plasticity
understanding of RNA export is partly due to a lack of of gene expression at the level of NPC export. To fully
faithful in vitro systems, which are difficult to establish understand the multifaceted behaviour of RNAs as
owing to inherent problems in coupling RNA transcrip­ they prepare to transit the nuclear pore will therefore
tion and processing to export. Although a few aspects require the integration of systems biology approaches
of RNA export have been reconstituted, we have not with detailed mechanistic studies.

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interactions with transcription factor IIH components. adaptor for Crm1–RanGTP‑mediated 60S subunit Acknowledgements
Mol. Biol. Cell 14, 1664–1676 (2003). export. We would like to acknowledge critical reading of the manu-
132. Lund, M. K. & Guthrie, C. The DEAD-box protein 148. Thomas, F. & Kutay, U. Biogenesis and nuclear export script by the members of the Hurt laboratory and by
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133. Weirich, C. S. et al. Activation of the DExD/H-box 149. Trotta, C. R., Lund, E., Kahan, L., Johnson, A. W. & The authors declare no competing financial interests.
protein Dbp5 by the nuclear-pore protein Gle1 and Dahlberg, J. E. Coordinated nuclear export of 60S
its coactivator InsP6 is required for mRNA export. ribosomal subunits and NMD3 in vertebrates. DATABASES
Nature Cell Biol. 8, 668–676 (2006). EMBO J. 22, 2841–2851 (2003). UniProtKB: http://ca.expasy.org/sprot
134. Alcazar-Roman, A. R., Tran, E. J., Guo, S. & Wente, S. R. 150. Nissan, T. A., Baßler, J., Petfalski, E., Tollervey, D. & CRM1 | Dbp5 | DGCR8 | Drosha | eEF1α | exportin-t |
Inositol hexakisphosphate and Gle1 activate the Hurt, E. C. 60S pre-ribosome formation viewed from exportin-5 | Los1 | Mex67 | Mlp1 | Mlp2 | Mtr2 | Nmd3 | Npl3 |
DEAD-box protein Dbp5 for nuclear mRNA export. assembly in the nucleolus until export to the Yra1
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References 133 and 134 show that Dbp5 is 151. West, M., Hedges, J. B., Chen, A. & Johnson, A. W. FURTHER INFORMATION
selectively activated on the cytoplasmic side of the Defining the order in which Nmd3p and Rpl10p load Ed Hurt’s homepage:
NPC to catalyse a terminal step of mRNA export. onto nascent 60S ribosomal subunits. Mol. Cell. Biol. http://www.uni-heidelberg.de/zentral/bzh
135. York, J. D., Odom, A. R., Murphy, R., Ives, E. B. & 25, 3802–3813 (2005).
Wente, S. R. A phospholipase C-dependent inositol 152. Hedges, J., West, M. & Johnson, A. W. Release of the SUPPLEMENTARY INFORMATION
polyphosphate kinase pathway required for efficient export adapter, Nmd3p, from the 60S ribosomal See online article: S1 (table)
messenger RNA export. Science 285, 96–100 subunit requires Rpl10p and the cytoplasmic GTPase All links are active in the online pdf
(1999). Lsg1p. EMBO J. 24, 567–579 (2005).

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