Scribd
Upload
5 views21 pages

Eukaryotic DNA Replication Mechanisms

Eukaryotic DNA replication is more complex than prokaryotes due to DNA being packaged in chromatin. It requires histones to dissociate and reassociate during replication. DNA polymerases α, δ, ε, γ, helicase, topoisomerase, primase, ligase, SSB proteins, dNTPs, Mg2+, and ATP are involved. DNA polymerase α initiates replication at origins of replication (ARS) by synthesizing an RNA primer. Polymerase δ and ε then perform leading and lagging strand DNA synthesis. Proliferating cell nuclear antigen (PCNA) aids polymerase processivity. Replication factor C loads PCNA onto DNA.

Uploaded by

anjali
Copyright
© All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
5 views21 pages

Eukaryotic DNA Replication Mechanisms

Eukaryotic DNA replication is more complex than prokaryotes due to DNA being packaged in chromatin. It requires histones to dissociate and reassociate during replication. DNA polymerases α, δ, ε, γ, helicase, topoisomerase, primase, ligase, SSB proteins, dNTPs, Mg2+, and ATP are involved. DNA polymerase α initiates replication at origins of replication (ARS) by synthesizing an RNA primer. Polymerase δ and ε then perform leading and lagging strand DNA synthesis. Proliferating cell nuclear antigen (PCNA) aids polymerase processivity. Replication factor C loads PCNA onto DNA.

Uploaded by

anjali
Copyright
© All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd

Eukaryotic DNA replication

• More complex than prokaryotes


• DNA in chromatin form
• Needs dissociation and reassociation of
histones
• Every time nucleosome has to dissembled
• Rate of synthesis – 50 nucleotides/sec
Proteins involved in replication
• DNA Polymerases α,δ,ε,γ.
• Helicase
• Topoisomerase
• Primase
• Ligase
• SSB Proteins
• dNTPs, Mg+2, & ATP
DNApolymeraseAlpha

• - Located in nucleus
• - Catalysis the initiation of replication on both
leading and lagging strand synthesis
• - Tetramer – 4 subunits POLA 1 (catalytic) POLA 1
• (regulatory) POLA3 ,4 (Primase)
• - larger subunit - 5´-3´ polymerization activity
• -Two smaller subunit – primase activity
• - one subunit – assist in other three
• subunits
• - RNA primer 5-15 nucleotides are subsequently
• extended by DNAPol α.
DNApolymerase Delta

• - Located in nucleus
• - Catalyzes the synthesis of lagging strand
• - Having four subunits – POLD 1,2,3,4
• - larger subunits catalyzes 5´-3´ polymerization
• activity
• - Smaller subunits catalyzes 3´-5´ exonuclease
• activity (proof reading activity)
• - High processivity when interacting with PCNA
• (Proliferating cell nuclear antigen).
PCNA

• - Molecular weight 25,000;


• PCNA is important for both DNAsynthesis and DNArepair
• Multimeric protein - Found in large amount in nuclei of
proliferating cells. - Act as “clamp” to keep DNA pol δ from
dissociating off the leading DNA strand.
• “Clamp” consist of 3 PCNA molecules each containing two
topologically identical domains that are tightly associated to form
closed ring. - PCNA helps hold DNA polymerase epsilon (Pol ε) to
DNA.
• DNA pol δ improves fidelity of replication by a factor of 102 due to
its proof reading action. It contributes in limiting the rates of overall
error to 10-9 to 10-12. - DNAPol δ is also associated with helicase
activity.
DNA polymerase Epsilon - Є
• located in nucleus
• Having four subunits – POLE 1, (Catalytic) 2,3,4
• (subunits)
• associated with - 5´- 3´ polymerization activity
• 5’- 3’ exonuclease activity (to remove RNAprimer)
• 3’- 5’ exonuclease activity (to proof read)
• DNA pol Є catalyzes the repair mechanism and catalyzes the removal of
primer and filing the primer gap in Okazaki fragments.
• Replicating factor A/ Replicating protein A (RPA/RFA)
• RPA/ RFA are similar to single strand binding protein. They bind to SS
DNA and prevent the reannealing of parental DNA.
Replication factor C (RFC)

• RFC also called as clamp loader or matchmaker.


• RFC assist in DNA pol δ to form clampbetween
DNA and PCNA.
• RFC also plays important role in setting up a
link between DNA pol δ and DNA pol α, so that
the
• leading strand synthesis and lagging strand
synthesis
• in eukaryotes can take place simultaneously.
ARS (Autonomously Replicating Sequences

• In eukaryotes the DNA replication is initiated at


• specific site known as ARS
• (Autonomously Replication Sequences) or replicators.
• ARS (Origin of chromosome in eukaryotes) contains
• - A central core sequence which contains highly
• conserved 11 bp sequence (AT rich sequence)
• - Flanking sequences. ARS – is 100- 150 long (generally it
span about 150 bp)
• There are multiple origins in eukaryotes
• In yeast 400
• Human chromosome contains average 100 ARS
ARS
• Consists of ORE and DUE

• ORE – 11 basepair region

• Binds to DNA pol, helicase, rcf forms ORC

• DUE – adjacent to ORE 80 bp region

• ORC - (DNA pol α, DNA pol δ, RFC, PCNA,


RFA, SSB and helicase)
Formation of pre RC

You might also like