DNA and

Replication

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History
of DNA

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 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
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 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 material

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Griffith Experiment

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 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 bacteria!
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 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
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 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

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 DNA
Structure

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 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 together by
weak hydrogen bonds
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DNA Double Helix
“Rungs of ladder”

Nitrogenous
Base (A,T,G or C)

“Legs of ladder”

Phosphate &
Sugar Backbone

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 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
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 DNA
• Stands for
Deoxyribonucleic acid
• Made up of subunits
called nucleotides
• Nucleotide made of:
1. Phosphate group
2. 5-carbon sugar
3. Nitrogenous base
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 DNA Nucleotide
Phosphate
Group

O
5
O=P-O CH2
O
O
N
Nitrogenous base
C4 C
1
(A, G, C, or T)
Sugar
(deoxyribose)
C C
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3 2
 Pentose Sugar
• Carbons are numbered clockwise
1’ to 5’ 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 copyright cmassengale P
18
 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)
Cytosine (C) T or 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,
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
Replication

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

5’ Parental DNA Molecule Replication

Fork
3’
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5’
 DNA Replication
• As the 2 DNA strands open at
the origin, Replication Bubbles
form
• Prokaryotes (bacteria) have a
single bubble
• Eukaryotic chromosomes have
MANY bubbles
Bubbles Bubbles

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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 untwisted

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 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

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 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

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 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
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 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
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 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 copyright cmassengale P
36
 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

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 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’ copyright cmassengale 5’
38
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
Replication Point of Origin
Fork

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 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
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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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