DNA REPLICATION

1. DNA Replication initiates from a single origin.

2. Replication Machinery.
3. Events at replication fork.
4. Termination of replication .
 Enzymes involved in
Initiation or Activation
are :

 Initiator protein Dna A.

 Dna B / Helicase enzyme.
 DNA gyrase.
 SSB proetins.
 DNA C : Helicase loader
 INITIATION OF DNA REPLICATION.

 Synthesis of DNA occurs at Replication fork .A

unit of replication with one origin is called
Replicon.
 DNA Replication begins at a single point called
Origin of Replication (Ori site).
 The initiator Protein/Enzyme Dna A is
responsible for triggering DNA replication it
binds with ori site and then it recruits Enzyme
Dna B helicase or helicase.
 Dna B /Helicase enzyme will break hydrogen bonds
holding two strands of double helix with the aid of
topoisomerase such as DNA gyrase.
 DNAgyrase relives supercoilling of DNA produced
as DNA strands are separated by helcase and DNA
gyrase also separates daughter molecules in final
stage of replication.
 Single stranded binding proteins (SSBs) keeps the
strands apart once they have been separated by
helicase.
Enzymes involve in
Main replication
machinery are :

1.Primase.
2.DNA polymerase-
III holoenzyme.
3.DNA polymerase I.
4.Ribonuclease H.
5.DNA ligase.
Replication machinery
 After the unwinding of DNA double
helix Primase enzyme synthesis
RNA primers as needed.
 DNA Polymerase enzyme catalyze
the synthesis of DNA.
 It has two limitations :
1. It needs primer for initiation.
2. It moves from 5’ to 3’.
 DNA polymeraseIII Holoenzyme

 It is multifunctional enzyme compose of 10

different proteins.
 It has 2 core enzyme and each binds one strand of
DNA and responsible for catalyzing DNA
synthesis and Proof reading.
 β-clamp tethers a core enzyme to DNA.
 Clamp loader at the centre is responsible for
loading β-clamp onto DNA
 Tau holds the holoenzyme together.
• LEADING STRAND :
it requires only one primer and DNA polymerase
enzymes synthesize DNA in the 5’ to 3’direction
according to base pairing rules and as it synthesis new
DNA strand continuously so that strand is named as
leading strand.
1. Lagging strand :
 lagging strand grows discontinuously by Forming
okazaki fragments. It requires multiple primers .
 Holoenzyme must discard old β-clamp And load new
and tether the template to core enzyme with each new
round of okazaki fragment synthesis.
• After most of lagging strand
has been synthesized by the
formation of Okazaki
fragments, DNA polymerase
I removes the RNA primers.

• Finally, the Okazaki

fragments are joined by the
enzyme DNA Ligase, which
forms a phosphodiester bond
between 3’-OH of the growing
strand and the 5’-phosphate of
an Okazaki fragment.
Termination:-
• In E.coli Replication stops at when replisome reach to
termination site (ter).
• A protein called tus (Termination utilisation sequence)
binds to ter site (tus-ter complex ) and than halts
progress of replication fork.
• In some bacteria Termination occurs spontaneously
when the fork meets .
• Recombinase enzyme catalyse an intermolecular
crossing over that separate the two chromosome.
• E.coli completes replication within 38 minutes i.e.
2000bp/s.