DNA Replication

AP Biology

2007-2008

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Synthesis Phase (S phase) S phase during interphase of the

cell cycle Nucleus of eukaryotes

DNA replication takes place in the S phase.

phase

G1

interphase

G2

-prophase -metaphase -anaphase -telophase

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Mitosis

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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Double helix structure of DNA

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Directionality of DNA
PO4

nucleotide

N base 5 CH2 O 4 3
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ribose
2

OH

The DNA backbone Putting the DNA

backbone together

5
PO4
5 CH2

base O
1
2

refer to the 3 and 5 ends of the DNA

the last trailing carbon

C
3

O O P O O 5 CH2
4
3

base O
1 2

OH
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Anti-parallel strands Nucleotides in DNA

backbone are bonded from phosphate to sugar between 3 & 5 carbons
DNA molecule has direction complementary strand runs in opposite direction

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Bonding in DNA
5
covalent phosphodiester bonds hydrogen bonds

.strong or weak bonds? AP Biology the bonds fit the mechanism for copying DNA? How do

Base pairing in DNA Purines

adenine (A) guanine (G)

Pyrimidines
thymine (T) cytosine (C)

Pairing

A:T
2 bonds

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C:G
3 bonds

Copying DNA Replication of DNA

new strand is 1/2 parent template & 1/2 new DNA

Semi-conservative

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Semiconservative??? Every daughter DNA molecule has an

intact template strand and a newly replicated strand.

Want Proof?

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Fact!

In the simplest model of DNA replication, new

nucleotides would be simultaneously added, according to the rules of complementarily, on both strands of newly synthesized DNA at the replication fork as the DNA opens up. But a problem exists, created by DNAs antiparallel nature; the two strands of a DNA double helix run in opposite directions. Going in one direction on the duplex, for example, one strand is a 5 3 strand, whereas the other is a 3 5 strand.

All polymerase enzymes add nucleotides in only

the 5 3 direction.
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DNA Replication
Large team of enzymes coordinates replication

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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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Replication: 1st step Unwind DNA

helicase enzyme
unwinds part of DNA helix
helicase

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replication fork

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

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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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Replication: 2nd step Build daughter DNA

strand
add new complementary bases DNA polymerase III

DNA Polymerase III

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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
3
RNA Primer

Nucleotide

DNA Polymerase

Direction of Replication
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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
3
RNA Primer

Nucleotides

DNA Polymerase

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Energy of Replication
Where does energy for bonding usually come from?

energy

ATP TTP GTP CTP

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CMP TMP GMP AMP

modified nucleotide

Replication Adding bases

energy

DNA Polymerase III

energy

can only add nucleotides to 3 end of a growing DNA strand

need a starter

DNA Polymerase III DNA Polymerase III

energy

nucleotide to bond to

energy

strand only grows 53

DNA Polymerase III

3 5

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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
5

3
5 3 5
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DNA Polymerase

RNA Primer

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Lagging Strand

energy no energy to bond

energy

energy

energy energy

ligase
energy

energy

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Lagging Strand Segments Okazaki Fragments - series of

short segments on the lagging strand Must be joined together by an enzyme
RNA Primer

Okazaki Fragment

DNA Polymerase

5
3
Lagging Strand

3
5
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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

Lagging Strand

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Leading & Lagging strands

Limits of DNA polymerase III

can only build onto 3 end of an existing DNA strand

3 3 growing 3 replication fork 5

Lagging strand

5 3

ligase
Leading strand
3

Lagging strand

5 3

Okazaki fragments joined by ligase

welder enzyme

DNA polymerase III

Leading strand

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continuous synthesis

Replication fork / Replication bubble

3 5 5 3

DNA polymerase III

leading strand 3 5 3 5 3 5 5 3 lagging strand 5

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Starting DNA synthesis: RNA primers

Limits of DNA polymerase III

can only build onto 3 end of an existing DNA strand

3 3 growing 3 replication fork 5 5 3 5

5 3

DNA polymerase III

primase
RNA 5

RNA primer
built by primase serves as starter sequence AP for DNA polymerase III Biology

Replication fork
DNA polymerase III
DNA polymerase I

lagging strand
primase

5 3

ligase

Okazaki fragments

3 5

SSB

3
DNA polymerase III

helicase

5 3
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leading strand
direction of replication
SSB = single-stranded binding proteins

Any Questions??

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2007-2008