1

DNA replication
Semiconservative replication
1- Separation of DNA strand occur
2- The new strand synthesized that are complementary to parent strand
3- The new DNA contain 2 strands one from parents and the other is
new strand

Basic requirement for DNA replication

1- Substrate
4 nucleotide of DNA (ATP, GTP, CTP, TTP) deoxy nucleotide triphosphate
(d NTP) must present
2- Template (parent strand)
Parent strand are required to add complementary nucleotide on the new strand
3- RNA primer
Present in the new strand and its function is to receive the first new nucleotide that
added on the new strand
4- Enzyme : DNA polymerase , Pol I, II , III
Step of replication in prokaryotic (E. coli)
1- Origin of replication
Replication start at particular DNA consensus sequence (AT rich region) called
origin (ori),
Dna A &B& C protein bind to strand produce separation of 2 strand
2- Formation of replication fork
Replication fork represent the unwinding the DNA strand by the effect of helicase
(Dna B) made broken in hydrogen bond lead to separation in helix this need 2 ATP
for each base pair broken
 2

Helicaes helped by 2 enzyme

1- Topoisomerase
Prevent super coil (super twist) that if occur lead to prevent replication
2- single stranded binding protein (SSBP)
Bind to single stranded DNA prevent it from reunion
Formation of new complementary strand
Always new DNA strand grow from 5 to 3 direction
A)Leading strand
This strand grow from 5 to 3 indirection
Of origin of replication
This grow continuously and made by Pol III
B) Okazaki fragment (lagging strand) 3
This strand grow from 5 to 3 direction in opposite of
origin of replication
This grow discontinuously and made by Pol III
DNA Pol I remove RNA primer then the gap filled
by Pol I and any space is ligated by DNA ligase enzyme
Okazaki fragment: about 1000-2000 nucleotide

Synthesis of DNA include following main steps

1- RNA primer
RNA primer are required to receive new nucleotide that added on new strand
because has OH free
Primase (type of RNA polymerase) enzyme synthesize RNA primer
Pol I remove it after termination of replication
There are 1 primer on leading strand and many primer in lagging strand

2- Prof reading effect of DNA Pol III

Prof reading means ability to remove any wrong nucleotide added during synthesis
as pairing A=G must be A=T
3- DNA Pol II
Has role in repair of DNA
 3

Comment Role in replication

DNA helicase (dan Broke hydrogen bond separation of helix
B)
SSBP Bind to single stranded DNA prevent it
from reunion

Primase synthesize RNA primer

DNA Pol III Synthesis of leading strand
Okazaki fragment
DNA Pol I Remove primer
Fill the gap in lagging strand
Gyrase Make negative super coil (super twist)
(topoisomeraseII)
DNA ligase Join the 2 end of DNA need ATP
 4

Eukaryotic DNA replication

1- Initiation
Separation of DNA double helix occur at multiple sites multiple replication
fork , this help to decrease time of replication
Eukaryotic DNA polymerase
1- Pol α synthesis of RNA primer and lagging strand
2- Pol β DNA repair
3- Pol γ synthesis of mitochondrial DNA
4- Pol δ synthesis of leading strand
5- Pol ε (epsilon) removal of RNA primer

What are telomeres and telomerase?

Replication end problem solved by telomerase
Telomere is a repeating DNA sequence (TTAGGG) at the end of the body's
chromosomes. . Telomeres function by preventing chromosomes from fusion of their
ends.
With each cell division, some of the telomere is lost (usually 25-200 base pairs per
division). When the telomere becomes too short, the chromosome reaches a "critical
length" and can no longer replicate. This means that a cell becomes "old" (senescent)
and dies

Telomerase, is an enzyme made of protein and RNA subunits that elongates

chromosomes by adding TTAGGG sequences to the end chromosomes.
Because these somatic cells do not regularly use telomerase, they age. The result of
aging cells is an aging body. If telomerase is activated in a cell, the cell will continue
to grow and divide. lead to cancer
Telomerase activated in germ cell and cancer cell

Importance of telomere
1- It present at the end of chromosome form break in DNA help to stop cell
cycle .
2- Help regulation of some protein
 5

Telomerase is reverse transcriptase that add base complementary to telomer

T A
T A
A Reverse transcription U
G C
G C
G C

DNA RNA
Telomer Telomerase

DNA Transcription RNA

Reverse transcription (telomerase)

Difference between Eukaryotic and prokaryotic DNA replication

Eukaryotic DNA Prokaryotic DNA

replication replication

1- 1-Origin of replication Multiple One

1- 2-Replication Multiple 2
fork
2- 3-Primer DNA Pol α Primase
synthesis
3- 4-Size of primer Long Short
4- 5-Lagging strand DNA Pol α DNA Pol III
synthesis
5- 6-DNA repair DNA Pol β DNA Pol II
6- 7-Leading strand Pol δ Pol III
7- 8-Telomerase Present Absent