DNA

REPLICATION
 DNA REPLICATION

• The process of DNA duplication

• It follows several steps that involve
multiple proteins called replication enzymes and RNA.
• In eukaryotic cells, such as animal cells and plant cells,
DNA replication occurs in the S phase of
interphase during the cell cycle.
• DNA, found within the nucleus, must be replicated in
order to ensure that each new cell receives the correct
number of chromosomes.
• The process of DNA replication is vital for cell growth,
repair, and reproduction in organisms.
 ENZYMES INVOLVE IN DNA
REPLICATION
• DNA helicase - unwinds and separates double stranded DNA as it
moves along the DNA. It forms the replication fork by breaking
hydrogen bonds between nucleotide pairs in DNA.
• DNA primase - a type of RNA polymerase that generates RNA
primers. Primers are short RNA molecules that act as templates for
the starting point of DNA replication.
• DNA polymerases - synthesize new DNA molecules by adding
nucleotides to leading and lagging DNA strands.
• Topoisomerase or DNA Gyrase - unwinds and rewinds DNA strands
to prevent the DNA from becoming tangled or supercoiled.
• Exonucleases - group of enzymes that remove nucleotide bases
from the end of a DNA chain.
• DNA ligase - joins DNA fragments together by forming
phosphodiester bonds between nucleotides.
STEP 1: REPLICATION FORK
FORMATION
• An enzyme called
Helicase breaks the hydrogen
bonds between the bases of the
two antiparallel strands.
• The strands are initially split
apart in areas that are rich in A-T
base pairs (there are only two
bonds between Adenine and
Thymine) forming a replication
fork.
• DNA Gyrase (also called
Topoisomerase) relieves tension
that builds up as a result of
unwinding.
• Single strand binding proteins
(SSBs) help to stabilize the
single stranded DNA.
STEP 2: PRIMER BINDING
• RNA polymerase (also
known as RNA Primase)
synthesizes short RNA
nucleotides sequences
that act as primers
(starters).
• These essentially
provide a starting point
for DNA replication.
STEP 3: ELONGATION
• DNA Polymerase III can now
start synthesising the new
DNA strand using free DNA
nucleotides.
• However, DNA polymerase can
only read the original template
(parent strand) in the 3’ → 5’
direction (making DNA 5’ →
3’).
• This is not a problem on the
leading strand, because the
DNA polymerase can simply
continue to read along as the
two parent stands continue to
unzip.
STEP 3: ELONGATION
• On the lagging strand DNA
polymerase moves away
from the replication fork.
• As the strands continue to
unzip more DNA is exposed
and new RNA primers must
be added.
• As a result the lagging
strand is synthesized in
short bursts as DNA
polymerase synthesizes
DNA in-between each of the
RNA primers.
STEP: ELONGATION
• The newly synthesised lagging
strand now consists of both
RNA and DNA fragments.
• The DNA fragments are known
as Okazaki fragments, after a
Japanese scientist who noticed
that heating DNA during
replication, which separates
the strands, gave many small
fragments of DNA.
• From this he concluded that
one strand must be
synthesized in short bursts of
DNA.
 STEP 4: TERMINATION

• DNA Polymerase I now removes the RNA primers and

replaces them with DNA
 STEP 4: TERMINATION
• DNA Ligase joins the DNA fragments of the lagging
strand together to form one continuous length of
DNA.
 TELOMERES
• During DNA replication the enzymes
(polymerase) that duplicate the chromosome and
its DNA can't continue their duplication all the
way to the end of the chromosome.
• At the very ends of the DNA, are long non-coding
region of Repeats known as telomeres.
• Every time the DNA is replicated the telomeres
shorten slightly.
• It is believed that this may be the genetic basis
for the aging process.
 PROOF READING AND CORRECTION
• DNA replication occurs at a surprisingly fast rate.
• Despite this, errors are very rare; occurring at a rate of
approx. 1 in every 10,000,000,000 base pairs.
• This is much lower than the expected value of about 1
in every 100 bp.
• This is the result of a complex series of enzymes that
proof-read the new DNA strands and make corrections
where needed.
 SUMMARY
• DNA replication is the production of identical DNA helices from a
single double-stranded DNA molecule.
• Each molecule consists of a strand from the original molecule and a
newly formed strand. Prior to replication, the DNA uncoils and
strands separate.
• A replication fork is formed which serves as a template for
replication.
• Primers bind to the DNA and DNA polymerases add new nucleotide
sequences in the 5ʹ to 3ʹ direction.
• This addition is continuous in the leading strand and fragmented in
the lagging strand.
• Once elongation of the DNA strands is complete, the strands are
checked for errors, repairs are made, and telomere sequences are
added to the ends of the DNA.
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