Presented by: Muhammad Tariq Aziz

2022-mphil-1411

MECHANISM
z
OF
REPLICATION
OF DNA
 z Introduction
o DNA replication is a biological process that occur in all living organisms
and copies their exact DNA. It is the basis for biological inheritance.

o It is actually a reaction in which daughter DNAs are synthesized using

prenatal DNAs as the template.

o Transferring the genetic information to the descendent generation with

high fidelity.

o DNA replication is semiconservative means one stand remains same in

the next formed DNA.
 z
Component of replication

 DNA polymerase:
deoxynucleotide polymerization

 Helicase:
 processive unwinding of DNA

 Topoisomerase:
 relieve torsional strain that results from helicase-induced unwinding
 z

 RNA primase:
Initiate synthesis of RNA primers

 Single strand binding proteins:

prevent premature reannealing of double stranded DNA

 DNA ligase:
Seals the single strand nick between the nascent chain and Okazaki fragment on
lagging strand

 Telomerase:
finishes off the ends of DNA strands in Eukaryotes
 z
DNA replication process

 There are three stage of DNA replication

1) Initiation

2) Elongation

3) termination
z
Initiation

 The replication starts at the

particular point called ‘origion
of replication (or) ori-sitr’.

 The structure of the origion is

248bp long and AT- rich.
 z
Elongation
 dNTPs are continuously connected to the primer or the nascent DNA chain by
DNA-pol III.

 The core enzymes ( α, ἐ , ᶿ ) catalyze the synthesis of leading and lagging

strands, respectively.

 The nature of chain elongation is the series formation of the phosphodiester

bonds.

 Many DNA fragments are synthesized sequentially on the DNA template strand
having 5’-end. The DNA fragments are called Okazaki fragments.

 They are 1000-2000 nt long in prokaryotes and 100-150 nt long in eukaryotes.

z
 z
Termination
 The replication of E.coli is bidirectional from one origin and two replication
fork must meet at one point called ter at 32.

 All the primers must be removed and the fragments must be connected by
DNA-pol I and ligase.

Telomeres:

 In eukaryotic replication following removal of RNA primer from the 5’ end

of the lagging strand, a gap is left. This gap exposes DNA strand to attack of
5’ exonucleases.

 THIS PROBLEM IS OVERCOME BY TELOMERASES.

 z SUMMARY OF REPLICATION

 Unwinding: forms replication fork

 Primase: synthesizes RNA primer

 Continuous synthesis: leading strand

 Discontinuous synthesis: lagging strand( Okazaki fragments)

 Synthesis: 5’-3’ direction

 Primers removed, nick sealed

 Proof reading by DNA polymerase (reduce error 1 in 10,000 to 1 in 100 million

bases)

 Organized in to chromatin structure