Scribd
Upload
121 views3 pages

Dna Replication Notes

DNA replication is the process by which DNA copies itself for cell division. It occurs during S phase of the cell cycle when chromosomes are not visible. During replication, the two strands of the DNA double helix separate and each strand acts as a template for new complementary strands to be synthesized. This results in two new DNA molecules that each contain one original strand and one newly synthesized strand, a process known as semiconservative replication. The Meselson-Stahl experiment provided evidence for semiconservative replication by showing that newly synthesized DNA contains a mix of heavy and light nitrogen isotopes. Replication occurs via a replication fork that forms at the origin of replication and progresses in both directions.

Uploaded by

Yash Vardhan
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
121 views3 pages

Dna Replication Notes

DNA replication is the process by which DNA copies itself for cell division. It occurs during S phase of the cell cycle when chromosomes are not visible. During replication, the two strands of the DNA double helix separate and each strand acts as a template for new complementary strands to be synthesized. This results in two new DNA molecules that each contain one original strand and one newly synthesized strand, a process known as semiconservative replication. The Meselson-Stahl experiment provided evidence for semiconservative replication by showing that newly synthesized DNA contains a mix of heavy and light nitrogen isotopes. Replication occurs via a replication fork that forms at the origin of replication and progresses in both directions.

Uploaded by

Yash Vardhan
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
You are on page 1/ 3

DNA METABOLISM (REPLICATION OF DNA)

The copying process of DNA to produce additional DNA molecules is known as replication.
It occurs in nucleus durg S phase of cell cycle when chromosomes are not easily visible.

DNA replication is semiconservative


 Watson and crick suggested that during replication the 2 strands of DNA double helix uncoil and separate, each
strand of DNA serves as a template for the synthesis of the new complementary strand.
 Due to Complementarity a nitrogenous base of a nucleotide in one strand, pairs up with the nitrogenous base
from another different nucleotide of another strand non-covalently by H bonds.
 After the completion of replication 2 new daughter DNA strands are synthesized which are identical to the
parent strands.
 Thus each of the daughter DNA double helices contains one original or old strand plus one new strand this style
of replication is said to be Semiconservative replication.

The Meselson – Stahl experiment to prove semiconservative replication of DNA

Note – (Under optimum conditions E.coli divides every 20 minutes.)

a) They grew E.coli in a medium containing 15N (Heavy nitrogen provided by 15NH4Cl). E.coli cells were grown for
many generations in a medium containing only heavy N i.e. 15N (blue), so that all the N in their DNA was 15N.
This heavy DNA molecule can be distinguished from the normal DNA by centrifugation in CsCl density gradient. It
was shown by a single blue band.
b) Then the E.coli cells with heavy nitrogen were transferred to a medium containing only light N i.e. 14N (red),
c) After 20 minutes (one division) cell were isolated and after centrifugation of DNA it was found that DNA
was a hybrid of 15N and 14N. it was shown by a purple band.
d) After 40 minutes i.e. second division sample of DNA was centrifuged and 2 DNAs were of hybrid density 14N15N
and 2 were light 14N14N.
e) Hybrid density DNA had one strand with 15N parental strand and other newly synthesized DNA strand with 14
N.
f) Light DNA had both the strands with 14N.
g) This experiment proved that every newly formed DNA has one parental strand and one newly synthesized strand.

Diagram from book here - -----------


Replication fork and origin of replication
 During replication entire DNA chain doesn’t split at once, DNA strands separate at specific point’s called Origin
of replication or Ori from where separation slowly progresses to the other end.
 Unzipping of the dsDNA forms a Y-shaped structure is called Replication fork.

Leading strand and Lagging strand


 A new strand of DNA is always synthesized in the 5’→3’ direction.
 Because the two DNA strands are antiparallel only one daughter strand is formed as a continuous stretch in 5’→3’
direction. This strand is called leading strand.
 On the other parent strand, daughter strand is synthesized in form of short DNA segments in 5’→3’ direction this
daughter strand is called Lagging strand.
 Lagging strand is not synthesized in smaller pieces and these short DNA strands are called Okazaki fragments.

Major enzymes involved in DNA replication


1. Helicase = unzipping enzyme
2. DNA Polymerase = Builder
3. Primase = Primer addition
4. Ligase = Glue
Steps involved in the process of DNA replication
1. Opening up of the double helix
2. Formation of RNA Primer
3. Elongation
4. Removal of the primer
5. Joining of okazaki fragments
6. Proof reading DNA and damage repair
7. Nucleotide excision repair
1. Opening up of the double helix –
 Eukaryotic DNA is bound to the histones to form nucleosome. During initiation of DNA replication it is made
accessible to the proteins and enzymes involved in the process of replication.
 Replication of DNA begins at specific sites where specific nucleotide sequences are present. They are called
replication origin or “ori” site.
 It is rich in A=T base pairs
Now helicase enzyme binds at ori site and unwinds the DNA. Due to unwinding Y shaped replication fork is
formed.
 Single strand binding proteins bind to the both strands, protecting them from rewinding.
 Unwinding generates a torque which is relieved by topoisomerase.
2. Formation of RNA primer –
 DNA polymerase III enzyme is used to add new nucleotides but it can not initiate new strand synthesis itself, it
only add new nucleotides to the existing strand to a primer is needed.
 RNA polymerase forms a RNA primer which is a small RNA molecule.
 RNA primer is later removed and gaps are filled up by DNA polymerase I.
3. Elongation –
 DNA nucleotides are activated and converted from deoxyribonucleoside monophosphates to
deoxyribonucleoside triphosphate.
 DNA polymerase adds new nucleotides in 5’→3’ direction only.
 As two parent DNA strands are oriented in antiparallel directions, one strand is synthesized continuously in
5’→3’ direction this strand is called leading strand.
 Another strand is synthesized in small segments called okazaki fragments which are later connected. This strand
is known as lagging strand.
 When polymerase reaches the 5’ end of the lagging strand, enzyme DNA ligase attaches the fragments.
 Because the synthesis of leading strand is continuous and that of lagging strand is discontinuous the overall
replication of DNA is said to be semi discontinuous.
4. Removal of the Primer –
 The enzyme DNA polymerase I removes the RNA primer and fills the gaps.
 It also fills the gaps of okazaki fragments.
5. Joining of Okazaki fragments
 After the gaps between the okazaki fragments are filled, the enzyme DNA ligase joins the fragments to form
lagging strand.
6. Proof reading of DNA and Damage repair –
 During replication DNA Polymerase proofreads each nucleotide as soon as it is added.
 Upon finding the incorrectly paired nucleotide the polymerase removes it and resumes synthesis.
7. Nucleotide excision repair –
 Damaged segment of DNA is cut or excised by nuclease enzyme, gaps are filled by DNA polymerase and joined by
DNA ligase.
 DNA repair enzymes in our skin cells are continuously repairing genetic damage caused by UV rays of sunlight.

You might also like