DNA Replication

(in eukaryotes)
By –Aditi Patriya
 Introduction
 Copying genetic information for transmission to the next generation .
 Occurs in S phase of cell cycle .
 Begins with the unwinding of the double helix to expose the bases in each
strand of DNA .
 Each unpaired nucleotide will attract a complementary nucleotide from the
medium – will form base pairing via hydrogen bonding.
 Enzymes link the aligned nucleotides by phosphodiester bonds to form a
continuous strand.
 About…
Complementary base pairing produces semiconservative replication .
• Double helix unwinds .
• Each strand acts as template .
• Complementary base pairing ensures that T signals addition of A on new
strand, and G signals addition of C .
• Two daughter helices produced after replication.
 The mechanism of DNA replication
 Tightly controlled process : occurs at specific times during the cell cycle.

 Requires:
• a set of proteins and enzymes,
• and requires energy in the form of ATP.
Two basic steps:
• Initiation
• Elongation
• Termination
 Mechanism enzymes
 DNA polymerase II – the enzyme that extends the primer.(DNA repair)
 Pol III – produces new stands of complementary DNA . (synthesis of leading and
lagging strand)
 Pol I – fills in gaps between newly synthesized Okazaki segments.
 Additional enzymes/proteins –
i. DNA helicase (Dna B)– unwinds double helix .
ii. Single-stranded binding proteins – keep helix open.
iii. Primase – creates RNA primers to initiate synthesis .
iv. Ligase – welds together Okazaki fragments.(sticky enzyme)
 Continue enzymes…..
 RNAase (DNA poly. Alpha)– remove RNA strand form by primase.
 Dna A (seq. A)– recognise the initiation site and stick to site.
 Dna C – assist Dna B .
 Telomerase- repair DNA naturally (maintain integrity of DNA telomere).
 Topoisomerase – helps uncoiling the supercoil of DNA .
 Origins of Replication

• Replication proceeds in both directions

(bidirectionally) from hundreds or thousands of
origins of replication on each of the linear
eukaryotic chromosomes.
 Replication Initiation

 DNA origin (Ori C) of replication.

 Initiator proteins (DNA a) bind .
 Recruits DNA helicase .
 Opening of DNA strands .
Direction of synthesis 5’ to 3’ .
SSB protein binds decrease wiling in strand .
Replication Fork
 Replication Elongation
Primer sequence is added (initiate replication ).
Topoisomerase become active.
Leading strand is made continuously .
Lagging strand made in pieces ( Okazaki fragments).
DNA poly 1 works with lagging strand (gap filling ).
RNAase will remove RNA strand form by primase.
 Replication Termination
Telomerase comes in power .
Works at telomere site .
Joins nucleotides in space of primase which is removed.
Synthesize DNA using RNA template.
protect chromosome ends from being degraded.
Telomerase is over expressed in cancer cells.
 DNA Topoisomerases
 enzymes that help relax the DNA by nicking the strands –
 Releasing the twists .
 Then rejoining the DNA ends.
 Example is DNA gyrase
THE END
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