DNA and

Replication

1
History
of DNA

2
 History of DNA
• Early scientists thought
protein was the cell’s
hereditary material
because it was more
complex than DNA
• Proteins were composed
of 20 different amino
acids in long polypeptide
chains 3
 Transformation
• Fred Griffith worked with
virulent S and nonvirulent R
strain Pneumoccocus
bacteria
• He found that R strain could
become virulent when it
took in DNA from heat-killed
S strain
• Study suggested that DNA
was probably the genetic 4
Griffith Experiment

5
 History of DNA
• Chromosomes are
made of both DNA
and protein
• Experiments on
bacteriophage
viruses by Hershey
& Chase proved that
DNA was the cell’s
genetic material
Radioactive 32
P was injected into
6
bacteria!
Discovery of DNA
Structure
• Erwin Chargaff showed
the amounts of the four
bases on DNA ( A,T,C,G)
• In a body or somatic cell:
A = 30.3%
T = 30.3%
G = 19.5%
C = 19.9%
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 Chargaff’s Rule
• Adenine must pair with
Thymine
• Guanine must pair with
Cytosine
• The bases form weak
hydrogen bonds

T A G C
8
 DNA Structure
• Rosalind Franklin took
diffraction x-ray
photographs of DNA
crystals
• In the 1950’s, Watson
& Crick built the first
model of DNA using
Franklin’s x-rays
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Rosalind Franklin

10
 DNA
Structur
e

11
 DNA
• Two strands coiled
called a double helix
• Sides made of a
pentose sugar
Deoxyribose bonded
to phosphate (PO4)
groups by
phosphodiester bonds
• Center made of
nitrogen bases bonded
12
DNA Double Helix
“Rungs of ladder”

Nitrogenous
Base (A,T,G or C)

“Legs of ladder”

Phosphate &
Sugar Backbone

13
 Helix
• Most DNA has a right-
hand twist with 10 base
pairs in a complete turn
• Left twisted DNA is called
Z-DNA or southpaw DNA
• Hot spots occur where
right and left twisted
DNA meet producing
mutations
14
 DNA
• Stands for
Deoxyribonucleic acid
• Made up of subunits
called nucleotides
• Nucleotide made of:
1. Phosphate
group
2. 5-carbon sugar
3. Nitrogenous 15
 DNA Nucleotide
Phosphate
Group

O
5
O=P-O CH2
O
O
N
Nitrogenous base
C4 C (A, G, C, or T)
1

Sugar
(deoxyribose) 16
C
3
C
2
 Pentose Sugar
• Carbons are numbered
clockwise 1’
5
to 5’
CH2

C 4 C1
Sugar
(deoxyribose)
C3 C2
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 5
DNA
O 3

3
O
P 5 P
5
O
1 G C 3
2
4 4
2 1
3 5
O
P P
5
T A 3

O
5
P 3 P
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 Antiparallel
Strands
• One strand of
DNA goes
from 5’ to 3’
(sugars)
• The other
strand is
opposite in
direction
going 3’ to 5’
(sugars)
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 Nitrogenous
Bases
• Double ring PURINES
Adenine (A)
Guanine (G)
A or G

• Single ring
PYRIMIDINES
Thymine (T)
T or C
Cytosine (C)
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 Base-Pairings
• Purines only pair with
Pyrimidines
• Three hydrogen bonds
required to bond
Guanine & Cytosine
3 H-bonds

G C
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• Two hydrogen bonds
are required to bond
Adenine & Thymine

T A

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 Question:
• If there is 30%
Adenine, how
much Cytosine is
present?

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 Answer:
• There would be 20%
Cytosine
• Adenine (30%) =
Thymine (30%)
• Guanine (20%) =
Cytosine (20%)
• Therefore, 60% A-T
and 40% C-G
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 DNA
Replicati
on

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 Replication Facts
• DNA has to be copied
before a cell divides
• DNA is copied during
the S or synthesis phase
of interphase
• New cells will need
identical DNA strands
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 Synthesis Phase (S
phase)
• S phase during interphase of
the cell cycle
• Nucleus of eukaryotes
S
DNA replication takes phase
place in the S phase.
G1 interphase G2

Mitosis
-prophase
-metaphase
-anaphase
-telophase 27
 DNA Replication
• Begins at Origins of Replication
• Two strands open forming
Replication Forks (Y-shaped
region)
• New strands grow at the forks 3’

Parental DNA Molecule

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 DNA Replication
• Enzyme Helicase
unwinds and
separates the 2 DNA
strands by breaking
the weak hydrogen
bonds
• Single-Strand Binding
Proteins attach and
keep the 2 DNA
strands separated and 30
 DNA Replication
• Enzyme Topoisomerase
attaches to the 2 forks of
the bubble to relieve stress
on the DNA molecule as it
separates
Enzyme Enzyme

DNA

31
 DNA Replication
• Before new DNA strands
can form, there must be
RNA primers present to
start the addition of new
nucleotides
• Primase is the enzyme that
synthesizes the RNA
Primer
• DNA polymerase can then
add the new nucleotides 32
33
 DNA Replication
• DNA polymerase can only add
nucleotides to the 3’ end of
the DNA
• This causes the NEW strand to
be built in a 5’ to 3’ direction
5’ 3’

RNA
5’
DNA Polymerase Primer
Nucleotide

Direction of Replication 34
 Remember HOW the
Carbons Are Numbered!
Phosphate
Group

O 5
CH2
O=P-O
O O
N
Nitrogenous base
C
4 C
1
(A, G, C, or T)
Sugar
(deoxyribose)
C3 C2 35
 Remember the Strands are
5
O
Antiparallel
3

3
O
P 5 P
5
O
1 G C 3
2
4 4
2 1
3 5
O
P P
5
T A 3

O
5
P 3 P
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 Synthesis of the New
DNA Strands
• The Leading Strand is
synthesized as a single
strand from the point of
origin toward the opening
replication fork
5’ 3’
5’
RNA
Nucleotides DNA Polymerase Primer

37
 Synthesis of the New DNA
Strands
• The Lagging Strand is synthesized
discontinuously against overall
direction of replication
• This strand is made in MANY short
segments It is replicated from the
replication fork toward the origin

Leading Strand
5 3’
’
3’ 5’
DNA Polymerase RNA Primer
5’ 3’

3’ 5’
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Lagging Strand
 Lagging Strand
Segments
• Okazaki Fragments - series
of short segments on the
lagging strand
• Must be joined together by
an enzyme DNA
Okazaki Fragment Polymerase
RNA
Primer
5’ 3’

3’ 5’
Lagging Strand
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 Joining of Okazaki
Fragments
• The enzyme Ligase joins the
Okazaki fragments together
to make one strand

DNA ligase
Okazaki Fragment 1 Okazaki Fragment 2
5’ 3’

3’ Lagging Strand
5’

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 Replication of
Strands
Replicati Point of
on Fork Origin

41
 Proofreading New
DNA
• DNA polymerase initially
makes about 1 in 10,000 base
pairing errors
• Enzymes proofread and
correct these mistakes
• The new error rate for DNA
that has been proofread is 1
in 1 billion base pairing errors
42
Semiconservative Model
of Replication
• Idea presented by Watson & Crick
• The two strands of the parental
molecule separate, and each acts
as a template for a new
complementary strand
• New DNA consists of 1
PARENTAL (original) and 1
NEW strand of DNA
DNA Template

Parental DNA
New DNA

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 DNA Damage &
Repair
• Chemicals & ultraviolet radiation
damage the DNA in our body
cells
• Cells must continuously repair
DAMAGED DNA
• Excision repair occurs when any
of over 50 repair enzymes
remove damaged parts of DNA
• DNA polymerase and DNA ligase
replace and bond the new
nucleotides together
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 Question:
• What would be the
complementary DNA
strand for the
following DNA
sequence?

DNA 5’-CGTATG-3’
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 Answer:

DNA 5’-CGTATG-3’
DNA 3’-GCATAC-5’

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