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DNA Replication

The document discusses DNA replication including the semi-conservative model, proteins involved such as DNA polymerase and primase, and differences between leading and lagging strand replication.

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erenyorulmaz143
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Download as PDF, TXT or read online on Scribd
16 views

DNA Replication

The document discusses DNA replication including the semi-conservative model, proteins involved such as DNA polymerase and primase, and differences between leading and lagging strand replication.

Uploaded by

erenyorulmaz143
Copyright
© © All Rights Reserved
Available Formats
Download as PDF, TXT or read online on Scribd
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Learning Objectives

Student Can
➢ Explain the overall mechanism of DNA
Replication
➢ Classify the proteins used in
machinery
➢ Differentiate the replication process
in two strands
DNA Replication
➢ Replication is the process of copying DNA strands.
➢ At the end of replication, one of the old strands is protected
(stayed as it is) as the DNA replication is semi-conservative.
➢ Adenin pairs with Thymidine
in DNA and Uracil in RNA.

➢ Guanine pairs with Cytosine


in both DNA and RNA

➢ According to Chargaff Rule;


# of A = # of T
# of G = # of C

➢ A binds to T via 2 hydrogen


bonds.
➢ G binds to C via 3 hyrdogen
bonds.
DNA Replication Models

❖ In the semi-conservative model, the two parental strands separate


and each makes a copy of itself. After one round of replication, the
two daughter molecules each comprises one old and one new
strand.
❖ After two rounds, two of the DNA molecules consist only of new
material, while the other two contain one old and one new strand.
DNA Replication Models

❖In the conservative model, the parental molecule directs synthesis


of an entirely new double-stranded molecule, such that after one
round of replication, one molecule is conserved as two old strands.

❖In the dispersive model, material in the two parental strands is


distributed more or less randomly between two daughter
molecules.
➢ During the
polimerization of
nucleotides in
replication, free 3’ end
hydroxyl group(3’-OH)
is used as an
attachment point of
polymerase enzyme
The replication
process occurs at
the centers which
are called
replication forks.
➢ In leading strand,the replication ends without any
interruption. Only a single primer is enough to finish the
replication of this strand.
➢ In lagging strand, due to the capability of polymerase to
synthesize only in 5’-3’ direction, there are many primers to
be synthesized via primase.
➢ In both strands, replication occurs in 5’-3’ direction
➢ Because DNA polymerase
needs a free 3’-OH end to
polymerase new
nucleotides enzyme
Primase synthesizes an
RNA primer to provide for
this free 3’-OH end need.

➢ Polymerase begins
polymerisation at that
point.
In the presence of any mistakenly
bonded nucleotide during replication, the
exonuclease activity of polymerase
prevents any base-pairing formation.
The error rate falls to 1 in 1010 in the presence of polymerase activity plus
proofreading activity (exonuclease) and plus mismatch repair system
➢ Replication Initiation

Helicase as a first step is responsible


for unwinding DNA strands
❑ Single strand binding proteins are responsible for prevention of
reforming H bonds. They let the strands stay apart from each other
during the replication process.
➢ Topoisomerases are enyzmes responsible for solving supercoiled DNA
strands especially in GC-rich DNA regions.
➢ Topoisomerase I excises off one of the two strands whereas
topoisomerase II cuts both.
Topoisomerase
mode of action
For Leading Strand replication;
➢ First of all, the enzyme helicase unwinds the double strands.
➢ Single strand binding proteins(SSBP) bind to strand letting them stay apart
from each other to prevent reformation of H bonds that have already
broken.
➢ Primase enzyme synthesizes an RNA primer to provide a free 3’-OH group for
polymerase.
➢ Via binding to this free 3’-OH end,polymerase starts polymerisation of new
strand of the DNA(replication).
➢ As the replication ends,polymerase removes the RNA primer and synthesizes
DNA sequence instead of it.
➢ The remained nicks between adjacent (next to) nucleotides then are filled
with a bond, called phosphodiester bond, and the enzyme that performs such
function is called DNA ligase or phosphodiesterase.
For Lagging Strand replication;
➢ First of all, the enzyme helicase unwinds the double strands like in leading
strand.
➢ Single strand binding proteins(SSBP) bind to strand letting them stay apart
from each other to prevent reformation of H bonds that have already
broken.
➢ Primase enzyme synthesizes a number of RNA primers to provide a free 3’-
OH group for polymerase.
➢ Via binding to this free 3’-OH end, polymerase starts polymerisation of new
strand of the DNA(replication) in 5’-3’ direction.
➢ The obtained approximately 300 nucleotide long fragments are called
«OKAZAKI fragments»
➢ As the replication ends, polymerase removes the RNA primers and
synthesizes DNA sequences instead of it.
➢ The remained nicks between adjacent (next to) nucleotides then are filled
with a bond, called phosphodiester bond, and the enzyme that performs such
function is called DNA ligase or phosphodiesterase.

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