DNA Replication

Reviewer
 DNA is the
instruction manual
for a living thing. Each
time one of your cells
divides, your DNA is
copied. That way, each
new cell has its own
copy of the instruction
manual.
DNA is converted into protein through the

The Central Dogma

replication
 The central dogma
illustrates the flow
of genetic
information from
DNA to RNA to
protein.
4
 DNA Replication
• DNA replication is the process of
producing two identical copies of DNA
• DNA must be copied before cell
division occurs
• DNA is copied during the S or
synthesis phase of interphase in the
nucleus
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 6
Semi-conservative model
of DNA Replication
when a new double-stranded DNA molecule is formed

• one strand will be from the original template

molecule
• one strand will be newly synthesized
• Replication is semi-conservative (one strand
is old, one strand new)
Step 1

Step 3
Step 2
 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

5’ Replication
Fork
3’

5’
10
DNA Replication
Enzyme 1: Helicase - unwinds and separates the 2 DNA strands

Singe-stranded binding proteins (SSBP) - attach and keep the 2

DNA strands separated and untwisted (prevents DNA molecule from
closing!)

Enzyme 2: Topoisomerase - relaxes supercoiled chromosome to

make DNA more accessible for the initiation of replication
helicase gyrase

single-stranded binding
proteins
replication fork
 Question…
• What kind of bonds does Helicase break?
– Hydrogen bonds
Topoisomerase prevents the DNA from getting too
tightly coiled ahead of the replication fork.
DNA Replication
Primase
Enzyme 3: RNA primase - adds
small section of RNA (RNA
5’
primer) that initiates DNA 3’

replication. 5’

Why must this be done?

RNA Primer

Enzyme 4: DNA polymerase 3 –

builds new DNA strand but can
only add nucleotides to existing
strands of DNA
DNA Polymerase III

Take note: DNA polymerase 3 needs RNA primer

before it starts initiating synthesis!
 DNA Polymerase builds
the new strand of DNA
en in a 5’ to 3’ direction
by adding matching
nucleotides
DNA
Polymerase III Which means DNA Polymerase
3 will only nucleotides on the
3’ end.
 Problem…
• How can both strands of DNA be
replicated in a 5’- 3’ direction at the
same time they are antiparallel?
• Answer: leading and lagging strands
 Leading and Lagging Strands
Limits of DNA polymerase III
◆ can only build onto 3 end of an existing DNA strand

5
3
5
3 5 5
3

Lagging strand
3 ligase
growing 3
replication fork
Leading strand

✓
5
3 5

Lagging strand 3
DNA polymerase III
◆ Okazaki fragments

Enzyme 5: DNA Ligase – “spot

welder enzyme” seals DNA Leading strand
fragments together ◆ continuous synthesis
Synthesis of the New DNA Strands
• The Leading Strand is synthesized
continuously as a single strand
from the point of origin toward
the opening replication fork

5’ 3’
5’
RNA
Nucleotides DNA Polymerase Primer

19
 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’
20
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
21
 DNA replication on the lagging strand
RNA primer is added
◆ built by primase
◆ serves as starter sequence for DNA polymerase III

HOWEVER short segments called Okazaki fragments are made

because it can only go in a 5 →3 direction

5

3 5 3
5
3
3 5

growing 3 primase
replication fork DNA polymerase III
5

RNA 5

3
 Replacing RNA primers with DNA
DNA polymerase I
◆ removes sections of
RNA primer and DNA polymerase I
replaces with DNA 5

nucleotides 3

3
5 ligase
growing 3
replication fork
5

RNA 5

3

STRANDS ARE GLUED

TOGETHER BY DNA LIGASE
 SSBs
Helicase DNA
Polymerase III

Leading Strand

DNA DNA Ligase

Polymerase I
Primase DNA Pol
Primer III Lagging
Strand
 Replication fork
DNA
polymerase III lagging strand
DNA
polymerase I
3’
Okazaki primase
fragments 5’
5’ ligase
3’ 5’ SSB

3’ helicase

DNA
polymerase III
5’ leading strand
3’
direction of replication
SSB = single-stranded binding proteins
 Finishing DNA Replication
• Problem #1: There are still RNA
nucleotides in the DNA (primers)
• Solution = DNA Polymerase 1 cuts out
the RNA nucleotides and replaces them
with DNA
 STEPS IN DNA REPLICATION
1-C. Helicase breaks the hydrogen bonds that connects the
nitrogenous bases
2-A. Single stranded binding proteins keep the two strands of
the DNA separate
3. Topoisomerase prevents the DNA from getting too tightly
coiled ahead of the replication fork.
3-E. Primer binds to the DNA strand as a point of origin
4-D. DNA polymerase binds to the strand at the site of the
primer and begins adding new base pairs complementary to the
strand
5-F. Okazaki fragments on lagging strands are joined together
by ligase
6-B. The parent strand and its complementary DNA strand coils
into the familiar double helix shape
Original New Original New
G C C G
C G G C

Sample Base Pairing

A T T A
A T T A
T A A T
C G G C
G C C G
G C C G
T A A T