348 C h ro m o s o m e B a n d i n g

occur in about 1 in 500 births. The most important Further Reading

types are 45,t(13q;14q) and 45,t(14q;21q), as these can Connor JM and Ferguson-Smith MA (1997) Essential Medical
lead to unbalanced offspring with translocation tri- Genetics, 5th edn. Oxford: Blackwell Science.
somy 13 (Patau syndrome) and translocation trisomy
21 (Down syndrome), respectively. See also: Adjacent/Alternate Disjunction;
Insertional translocations may occur within a Down Syndrome; Idiogram; Isochromosome;
chromosome or between two chromosomes. Three Karyotype; Klinefelter Syndrome; Patau
breakpoints are required, two to provide the inter- Syndrome; Polyploidy; Prenatal Diagnosis;
stitial deletion of a chromosome fragment and one Triploidy; Trisomy 18; Turner Syndrome
to allow the insertion of the fragment into another site.
Segregation of the two translocation derivatives during
meiosis may lead to unbalanced offspring with either a
deletion or duplication of the inserted fragment.
Chromosome Banding
Most deletions and duplications are the result of A T Sumner
unbalanced translocations. Ring chromosomes arise Copyright ß 2001 Academic Press
when breaks occur at both ends of a chromosome, doi: 10.1006/rwgn.2001.0204
with reunion of the proximal ends and loss of the distal
telomeric fragments. They can be associated with sub-
stantial terminal deletions of DNA. Ring chromo- Definition and History
somes are a common feature of irradiated cells. They
Chromosome banding is the ``lengthwise variation in
may occur as part of a constitutional abnormality but
staining properties along a chromosome . . . normally
are seldom inherited unless very small. Some are
independent of any immediately obvious structural
unstable, and sister chromatid exchange within the
variation,'' and thus excludes patterns such as those
ring may lead to double-sized dicentric rings. The
seen on polytene chromosomes of Drosophila, which
instability of the dicentric ring may lead to further
have a morphological component. Although the first
changes and these may be associated with more exten-
observations of what could be called chromosome
sive phenotypic abnormality.
banding were made at the end of the nineteenth cen-
An isochromosome is a metacentric chromosome
tury, modern chromosome banding methods date
in which both arms are genetically identical (see Iso-
from 1968 and can be applied to chromosomes of a
chromosome). It most often arises by an isochromatid
wide variety of species with no more than slight modi-
break and fusion of the sister chromatids above the
fications. Following the introduction of Q-banding
centromere. Thus most isochromosomes are dicentric,
by Caspersson and his colleagues in 1968, Pardue
although only one centromere is active and the iso-
and Gall inadvertently produced differential staining
chromosome segregates normally during cell division.
of heterochromatin in their pioneering in situ hybri-
Human isochromosomes occur mostly as sex chromo-
dization studies, leading directly to C-banding, and in
some abnormalities and are particularly associated
1971 G-banding was discovered by several authors. R-
with Turner syndrome (see Turner Syndrome).
banding was also introduced in 1971. Over the
Inversions are intrachromosomal aberrations
next few years, many other banding techniques, too
which result from two breaks with inversion of the
numerous to mention individually, were introduced,
intervening segment through 1808. There are essen-
many of them using fluorochromes. Silver staining for
tially two types: pericentric inversions, in which the
nucleolus organizing regions (NORs) was introduced
centromere is included within the inversion; and para-
in 1975, methods to show chromosome replication
centric inversions in which the centromere is outside
were invented, and the use of autoimmune sera to
the inverted segment. An inversion alters the order of
label kinetochores immunocytochemically was dis-
gene loci within a chromosome and this in itself has no
covered.
phenotypic effect. However, inversions interfere with
synapsis of homologs during meiosis. An inversion
loop may form in order to achieve synapsis. Cross- Classification of Chromosome Bands
ing-over within the loop of a paracentric inversion
Four classes of bands can be recognized:
leads to dicentric and acentric recombinants. Cross-
ing-over within a pericentric inversion leads to mono- 1. Heterochromatic bands are demonstrated by C-
centric recombinants with duplication and deletion of banding techniques, as well as by various methods
the chromosome distal to the breakpoints of the inver- of fluorochrome staining, and correspond to clas-
sion. The closer the inversion breakpoints are to the sically defined constitutive heterochromatin, that
ends of the chromosome, the smaller the imbalance. is, regions of chromosomes that normally remain
 Chromosome Banding 349

condensed throughout interphase, and are gener- CREST labeling of kinetochores is an important
ally found as blocks around centromeres, and tool in identifying centromeres and in distinguishing
sometimes terminally or interstitially on chromo- active and inactive centromeres in dicentric chromo-
somes. Facultative heterochromatin, such as the somes, and has become invaluable in understanding
inactive X chromosome in female mammals, is not centromeric organization.
stained specifically by banding methods.
2. Euchromatic bands form a pattern of alternating
positively and negatively stained (or fluorescent) Functional and Structural Significance of
bands throughout the length of the chromo- Chromosome Bands
somes, and are demonstrated by methods such as Heterochromatin is widely believed to be functionless
G-banding, R-banding, Q-banding, and by certain (`junk DNA'), a view supported by the lack of obvious
fluorochromes. phenotypic effect of C-band polymorphisms in many
3. Nucleolus organizer regions are the segments of cases (see above), and by its content (in most cases) of
chromosomes that contain the genes for ribosomal highly repetitive DNA sequences that could certainly
RNA, and which give rise to the interphase not code for proteins, and which, it seems to many
nucleoli. They can be stained with silver (Ag± people, could have no other conceivable function.
NOR staining). Such views are almost certainly incorrect. In Dros-
4. Kinetochores are the centromeric structures ophila, a number of genes, as well as some nongenic
through which mitotic and meiotic chromosomes functions, have been localized to heterochromatin,
are attached to the spindle microtubules, and are and it could well be that when the heterochromatin
generally labeled using autoimmune CREST sera. of other organisms has been examined in the same
amount of detail, it will be found that these too have
various functions in their heterochromatin. In add-
Applications of Chromosome Banding ition, it has been suggested that centromeric hetero-
The most important application of banding is in the chromatin has an essential role in holding sister
identification of individual chromosomes. Euchro- chromatids together until the end of metaphase, and
matic banding techniques, especially G-banding, are ensuring their controlled separation at the beginning
ideal, but euchromatic bands are essentially restricted of anaphase.
to higher vertebrates. However, even in organisms Many differences have now been found between
that lack euchromatic bands, distinctive patterns of positive and negative euchromatic bands, most of
heterochromatic banding can often be used to distin- which are related to the fact that positive G-bands
guish between chromosomes. In humans, G-banding have relatively few genes, while negative ones are
is used to identify chromosome abnormalities and much richer in genes; in humans, approximately 80%
rearrangements in genetic diseases and cancers. Band- of the genes are in the negative G-bands, which form
ing is also valuable for the identification of chromo- only about half of the genome. The highest concen-
some rearrangements that have occurred in the course trations of genes are in the T-bands, a subset of
of evolution. R-bands (negative G-bands) that are found largely at
Polymorphisms of heterochromatic bands have the ends of chromosomes.
become a study in their own right, as well as being The reason for the division of chromosomes into
useful tools for distinguishing between paternal and gene-rich and gene-poor segments is not at all clear,
maternal homologs. There is no evidence that these but it is probably universal, and not restricted to
polymorphisms have any phenotypic effects in mammals. Evidence is accumulating that lower verte-
humans, but in maize there is a correlation between brates, invertebrates, and plants also show a nonuni-
growth rate and amount of heterochromatin, suggest- form distribution of genes on their chromosomes,
ing that in some cases, heterochromatin may have which appears to be correlated with patterns of early
phenotypic effects. and late replication. Chromosome banding, therefore,
Ag±NOR staining can be used not only to identify is not simply an invaluable method of identifying
the location of nucleolus organizers on chromosomes, chromosomes, but has also become very important
but also to assess their activity. In a species with multi- in drawing our attention to functional aspects of the
ple NORs, such as humans (who have five pairs) only longitudinal differentiation of chromosomes.
a proportion are stainable with silver, while in hybrids,
it often happens that only the NORs from one parent Further Reading
are active. Ag±staining of NORs in interphase nuclei Bickmore W and Craig J (1997) Chromosome Bands: Patterns in
also has prognostic value in various cancers. the Genome. Austin, TX: R. G. Landes.
350 C h ro m o s o m e B re a k

Sumner AT (1990) Chromosome Banding. London: Unwin uncapped chromosome. Chromosome breaks can
Hyman. also be made by a variety of agents external to the
cell. Ionizing (gamma or X-ray) radiation induces
See also: Centromere; Heterochromatin; breaks, as do a variety of chemicals. Alkylating agents
Kinetochore; Nucleolus (e.g., methyl methane sulfonate), base derivatives, aro-
matic amines, nitroso compounds, and heavy metals
can also cause chromosome breaks.
Chromosome Break There are two pathways to repair chromosome
breaks. Repair by homologous recombination uses
J Y Lee and T L Orr-Weaver the genetic information from the sister chromatid or
Copyright ß 2001 Academic Press homolog as the template for repair. If a sister chroma-
doi: 10.1006/rwgn.2001.0205 tid is available (i.e., after DNA replication and before
mitosis), the repair process should restore the chromo-
some to its original state. However, if repair is per-
A chromosome break is a break in the phosphodiester formed using the sequence of the homolog and the
backbone of DNA. Here chromosome break is under- homolog sequence is different, the repaired chromo-
stood to be a double-strand break in the DNA. some will carry a new sequence. This may lead to
The consequences of unwanted chromosome breaks loss of heterozygosity at that locus and the possibility
are severe. The resulting broken ends of DNA are of a detrimental phenotype. Repair by the nonhom-
recombinogenic, and this can lead to chromosome ologous end-joining (NHEJ) pathway does not use a
fusions, aneuploidy, or rearrangements such as inver- copy of the chromosome, but instead brings two
sions, translocations, and deletions. In certain plants pieces of broken DNA together and joins their ends.
and animals, a chromosome break can be the first stage While this process may restore the overall structure of
of a `breakage±fusion±bridge' cycle. The loose ends the DNA, it is usually not precise and deletions can
made by a chromosome break may fuse together and occur. Though proteins of both pathways are con-
form a bridge between two different chromatids or served throughout evolution, yeast preferentially use
chromosomes. The resulting dicentric chromosome is homologous recombination, while mammalian cells
unstable, and another chromosome break forms, lead- more frequently employ NHEJ.
ing to duplications and deletions. Mutations in repair genes may cause serious diffi-
In vivo, chromosome breaks are formed in several culties for an organism. In humans, there are several
normal cellular processes. During T and B cell differ- diseases in which genes in the NHEJ pathway are
entiation, a double-strand break is made during V(D)J mutated and patients are highly cancer-prone. In
recombination such that the different gene segments ataxia-telangiectasia-like disorder, the hMRE11 gene
are cut and joined together by the nonhomologous is mutated and in Nijmegen syndrome, NBS1 is
end-joining pathway (see below). During meiosis in altered. Both of these proteins are in the NHEJ repair
yeast, recombination is initiated by the formation of a pathway, along with the Ku proteins and DNA-
double-strand break by the enzyme Spo11. The Spo11 dependent protein kinase.
gene is conserved to mammals and is essential for
meiosis and synaptonemal complex formation in the See also: Chromatid; Chromosome
mouse, suggesting that formation of double-strand
breaks may be a general mechanism for initiating
meiotic recombination. The cut ends formed by the
breaks go on to find nonsister chromatids and exchange
Chromosome Bridge
sequences. J S Heslop-Harrison
Errors in cell metabolism can lead to chromosome Copyright ß 2001 Academic Press
breaks. Incomplete replication of the chromosomes doi: 10.1006/rwgn.2001.0206
can lead to difficulties when the sister chromatids are
segregated at mitosis. The pull of the spindles can
break off a part of a chromatid that does not have Chromosome and chromatid bridges occur at ana-
sister sequence because of unfinished replication. An phase of mitosis and meiosis when chromatids are
improper telomere `cap' of chromosomes can also lead not free to separate and form a bridge between the
to breaks. The telomere is a protective structure for two sets of segregating chromosomes. The chromatid
the cell, ensuring that no genetic information is lost at or chromosome forming the bridge usually breaks,
replication. An uncapped chromosome can be recom- leading to duplication of a segment in one daughter
binogenic and is more likely to fuse with another nucleus and deletion in the other. Bridges occur for