DNA Replication

Replication
The duplication of DNA which occurs
during the S phase of Interphase.
1 Strand 2 Complementary Strands
DNA Polymerase

 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

Features of DNA Replication

DNA replication is semiconservative
Each strand of template DNA is being copied.

DNA replication is bidirectional

Bidirectional replication involves two replication forks,
which move in opposite directions

DNA replication is semidiscontinuous

The leading strand copies continuously
The lagging strand copies in segments (Okazaki
fragments) which must be joined

Hydrogen bond
(H-bonds)

thymine
adenine
cytosine
guanine
phosphate
deoxyribose sugar

Sugar / phosphate strand

Nitrogenous base rung

DNA nucleotide
Deoxyribose sugar

phosphate

RNA nucleotide
ribose sugar

Nitrogenous base
(guanine)

phosphate

Nitrogenous base
(uracil)

Step 1: Hydrogen bonds

between complimentary
bases break
DNA unzips

Step 2: DNA strands

pull apart from each other

Step 3: DNA nucleotides in the cell

match up with each side of the
unzipped DNA
each unzipped strands forms a
template for a new strand

Step 4: Each old strand

forms a template for a
new strand
two identical DNA
molecules form

new strand, identical

sequence to the original

old (original) strand

MODELS OF DNA REPLICATION

(a) Hypothesis 1:

(b) Hypothesis 2:

(c) Hypothesis 3:

Semi-conservative
replication

Conservative replication

Dispersive replication

Intermediate molecule

Replication can be Uni- or Bidirectional

UNIDIRECTIONAL REPLICATION
Origin
BIDIRECTIONAL REPLICATION
3
5

5
3
5
3

Origin
3
5

MECHANISM OF DNA REPLICATION

summary of DNA replication
know structure, directionality, enzymes, functions

Proteins at the replication fork in E. coli

Rep protein (helicase)

3
5

pol III

5
3

Primasome

DNA ligase

C B
Single-strand
binding protein
(SSB)

pol III

DNA gyrase - this is a topoisomerase II, which

breaks and reseals double-stranded DNA to introduce
negative supercoils ahead of the fork

pol I

Enzymes in DNA replication

Helicase unwinds
parental double helix

DNA polymerase
binds nucleotides
to form new strands

Binding proteins
stabilize separate
strands

Exonuclease removes
RNA primer and inserts
the correct bases

Primase adds
short primer
to template strand

Ligase joins Okazaki

fragments and seals
other nicks in sugarphosphate backbone

Replication
3
3

5
3
5

Helicase protein binds to DNA sequences called

origins and unwinds DNA strands.
Binding proteins prevent single strands from rewinding.
Primase protein makes a short segment of RNA
complementary to the DNA, a primer.

Replication
Overall direction
of replication

3
3

5
3
5

3
5

DNA polymerase enzyme adds DNA nucleotides

to the RNA primer.

Replication
Overall direction
of replication

3
5

5
3
5

3
5

DNA polymerase enzyme adds DNA nucleotides

to the RNA primer.
DNA polymerase proofreads bases added and
replaces incorrect nucleotides.

Replication
Overall direction
of replication

3
3

5
3
5

Leading strand synthesis continues in a

5 to 3 direction.

3
5

Replication
Overall direction
of replication

3
3

5
Okazaki fragment

3
5

3 5

3
5

Leading strand synthesis continues in a

5 to 3 direction.
Discontinuous synthesis produces 5 to 3 DNA
segments called Okazaki fragments.

Replication
Overall direction
of replication

3
3

5
Okazaki fragment

3
5

3 5

3
5

Leading strand synthesis continues in a

5 to 3 direction.
Discontinuous synthesis produces 5 to 3 DNA
segments called Okazaki fragments.

Replication
3
5

3
5
3
5

3 5

35

3
5

Leading strand synthesis continues in a

5 to 3 direction.
Discontinuous synthesis produces 5 to 3 DNA
segments called Okazaki fragments.

Replication
3
5

3
5
3
5

35

35

3
5

Leading strand synthesis continues in a

5 to 3 direction.
Discontinuous synthesis produces 5 to 3 DNA
segments called Okazaki fragments.

Replication
3
5

3
5
3
5

35

35

3
5

Exonuclease enzymes remove RNA primers.

Replication
3
3
5
3
5

35

3
5

Exonuclease enzymes remove RNA primers.

Ligase forms bonds between sugar-phosphate
backbone.

Replication
5
3

3
5
3

3
5

5
3

5
5

5
3

3
5
3

5
5

5
3

3
3
5

5
3 5

3
3
5

5
3

3
5

5
3
5

Components of the replication apparatus

dnaA
Primasome
dnaB
dnaC
dnaG
DNA gyrase
Rep protein
SSB
DNA pol III
DNA pol I
DNA ligase

binds to origin DNA sequence

helicase (unwinds DNA at origin)
binds dnaB
primase (synthesizes RNA primer)
introduces negative supercoils ahead
of the replication fork
helicase (unwinds DNA at fork)
binds to single-stranded DNA
primary replicating polymerase
removes primer and fills gap
seals gap by forming 3, 5-phosphodiester bond