DNA Replication Process in Living Organisms
DNA replication
is the process of producing two identical copies from one
original
DNA
molecule. This biological process occurs in
all
living organisms . It
is the basis for
biological
inheritance
. DNA is composed from two strands and each
strand of the original DNA molecule serves as
template
for
the production of the complementary strand, a process
referred to as
semiconservative replication
and the
process followed by
proofreading
or error-checking
mechanisms to ensure correct reading to the genetic
code,. like all biological polymerization
processes(Transcription and Translation
), the process
involve 3 stages :
1- initiation
2-elongation and 3- termination
.
Steps of DNA replication
1- Initiation: the process require
replicator
and
initiator
protein
.
For a
cell to divide
, it must first replicate its DNA.
This process is initiated at particular points in the DNA,
known as replicator (200-300 bp) which contain specific
area called "
origin of replication ori c)
", which will
opened by
initiator proteins
.
Sequences opened by initiator proteins tend to be "AT-
rich" (rich in adenine and thymine bases), because A-T
base pairs have two hydrogen bonds (rather than the
three bond in a C-G pair).
Once the origin has been recognized, the initiators
proteins (
origin recognition complex
)
start forming
the
pre-replication complex
, which unwind the double-
stranded DNA by helicase enzyme.
All known DNA replication systems require a free
3'
hydroxyl
group before synthesis can be initiated. use
a
primase enzyme
to synthesize a short RNA primer(10-20 bp)
with a free 3′ OH group which is subsequently elongated by a
DNA polymerase in this mechanism,
In eukaryotes, primase is produced by Pol α DNA polymerase
helicases, which break the hydrogen bonds holding the two DNA
strands together. These two strands serve as the template for the
leading and lagging strands, which will be created as DNA
polymerase matches complementary nucleotides to the
templates; the templates may be properly referred to as the
leading strand template and the lagging strand templates.
Single strand binding proteins (SSBPs)
bind to the single
strands of DNA near the replication fork to prevent the
ssDNA strands from winding back into a double helix,
thus
maintaining the strand separation.
2- Elongation step
DNA is always synthesized in the 5' to 3' direction.
Since the leading and lagging strand templates are
oriented in opposite directions at the replication fork,
a major issue is how to achieve synthesis of nascent
(new) lagging strand DNA, whose direction of
synthesis is opposite to the direction of the growing
replication fork.
A new DNA strand is always synthesized in a 5’ to 3’ manner, thus
the replication of both the strands goes in two different ways.
1- Leading strand .
A leading strand is the strand which is run from 5’-3’direction or
the direction the same as the replication fork movement. It is
synthesized continuously; there are no breaks in-between. This
strand is formed as nucleotides are continuously added to the 3’
end of the strand after polymerase reads the original DNA
template . Only one primer will require here .no Okazaki
fragment will formed.
2 DNA polymerase
molecules are required for polymerization
the two strands
the
leading
and
lagging
strands are synthesized by
Pol ε
(epsilon)
and
Pol δ(
delta)
,
respectively.
Recently
replication termination involves a switch from Pol ε to Pol
δ ; thus, the commonly accepted model is that Pol δ performs
initiation and termination on both strands, as well as the
synthesis of Okazaki fragments, whereas Pol ε elongates only the
leading strand.
1-The leading strand receives one RNA primer while the
lagging strand receives several
2- The leading strand is continuously extended from the
primer by a high
processivity
, replicative DNA polymerase,
while the lagging strand is extended discontinuously from
each primer, forming Okazaki fragments As DNA synthesis
continues, the original DNA strands continue to unwind on
each side of the bubble, forming
a replication fork
with two
prongs.
β
Clamp proteins : it
form a sliding clamp around DNA,
helping the DNA polymerase maintain contact with its
template, thereby assisting with processivity. The inner face
of the clamp enables DNA to be threaded through it.
Once the polymerase reaches the end of the template or
detects double-stranded DNA, the sliding clamp undergoes
a conformational change that releases the DNA
polymerase.
2- Lagging strand :it
DNA polymerase can extend the RNA
primer, adding nucleotides one by one that are complementary to the
template strand. ( example:
A
in the template strand is complement
to
T
in new growing strand, and
G
in the template strand is
complement to
C
in new growing strand).
A DNA polymerase
delta (δ)
extends the primed segments,
forming Okazaki fragments.
DNA polymerase will add nucleotides in the 5' to 3'
direction; however, one of the parent strands(lagging) of
DNA is 3' to 5' while the other (leading) is 5' to 3'.
To solve this problem, replication occurs in opposite
directions. lagging strand run away from the replication
fork, and synthesized a series of short fragments known as
Okazaki fragments, consequently requiring many primers.
The RNA primers of Okazaki fragments are degraded by
Rnase H.
Types of DNA polymerase in Eukaryotic cell
The DNA polymerases of eukaryotes are
in general less understandable than the DNA
polymerases of prokaryotes. Eukaryotic cells have
FIVE
polymerases:
four
major
nuclear
DNA polymerases:
DNA polymerase alpha (Pol α), DNA polymerase delta (Pol δ) and
DNA polymerase epsilon (Pol ε), DNA polymerase beta ( poly
β
),
and
one
found in
mitochondria
:
DNA polymerase Gamma (Poly
γ
).
Plant chloroplasts and
Plant chloroplasts and
mitochondria
also contain their own DNA polymerase that appears to be similar
also contain their own DNA polymerase that appears to be similar
to
to
γ
γ
1-
Polymerase
alpha ( Pol
α
)
:
it is the only enzyme has
primase activity
beside DNA
polymerase,
it is the only enzyme has primase activity beside DNA polymerase so
it is the only enzyme has primase activity beside DNA polymerase so
it is self- primed it will form short primer 12-20 nts called the initiator RNA iRNA
it is self- primed it will form short primer 12-20 nts called the initiator RNA iRNA
2-Pol
β
Beta
polymerase
:
excision repair
and it is not highly active and is not very
processive.
3-Pol
γ
Gamma
polymerase
: polymerization the
mitochondrial DNA
beside
repairing
by its
exonuclease activity 3́→5́ in mammals.
4-Pol
δ
delta
and
5-
ε
epsilon
polymerase
:polymerization lagging (
δ
)and leading (
ε
) strand
respectively 5 ́→3́. In
eukaryotes
.
Termination in Eukaryotic cell
Primer removal in eukaryotes is performed by
RNase I
that removes
all the primer leaving only one nucleotide in the junction between 2
nucleotide and the remained one will removed by
FenI enzyme.
a single nick on the leading strand and several nicks on the lagging
strand can be found.
Ligase
works to fill these nicks in, thus
completing the newly replicated DNA molecule.
Eukaryote cell initiate DNA replication at multiple points in the
chromosome, so replication forks meet and terminate at many points
in the chromosome; Because eukaryotes have linear chromosomes,
DNA replication is unable to reach the very end of the chromosomes,
but ends at the
Telomere
region of repetitive DNA close to the end.
This shortens the telomere of the daughter DNA strand. Shortening
of the telomeres is a normal process in
Somatic
cells.
Within the
Germ cell
line, which passes DNA to the next
generation,
Telomerase
extends the repetitive sequences of the
telomere region
in the end of the replication the DNA will warp around the basic
histones to form the chromatin
The problem is that
semiconservative DNA
replication requires an RNA
primer for initiation, and the
lagging strand requires one
primer to start each Okazaki
fragment. Even if an RNA
primer can be placed at the very
39 end of the chromosome
replicated as the lagging strand,
this primer eventually will be
lost because no distal Okazaki
fragment can be extended
through it. The loss of 10 or 12
nucleotides (or more, if the last
primer is not positioned at the
very end) at each cell division
should pose major problems for
long-lived multicellular
eukaryotes.
DNA replication is a fundamental biological process where an original DNA molecule produces two identical copies. This process involves initiation, elongation, and termination stages, utilizing replicator and initiator proteins. The DNA is unwound and replicated with the help of enzymes like helicase and DNA polymerase. Elongation occurs in the 5' to 3' direction, with the leading and lagging strands synthesized in opposite directions. Various mechanisms ensure accurate replication, such as primase synthesis of RNA primers, DNA polymerase matching complementary nucleotides, and single-strand binding proteins preventing helix winding.
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Presentation Transcript
DNA replication is the process of producing two identical copies from one original DNA molecule. This biological process occurs in all living organisms . It is the basis for biological inheritance. DNA is composed from two strands and each strand of the original DNA molecule serves as template for the production of the complementary strand, a process referred to as semiconservative replication and the process followed by proofreading or error-checking mechanisms to ensure correct reading to the genetic code,. like all processes(Transcription and Translation), the process involve 3 stages : 1- initiation 2-elongation and 3- termination. biological polymerization
Steps of DNA replication 1- Initiation: the process require replicator and initiator protein. For a cell to divide, it must first replicate its DNA. This process is initiated at particular points in the DNA, known as replicator (200-300 bp) which contain specific area called "origin of replication ori c) ", which will opened by initiator proteins. Sequences opened by initiator proteins tend to be "AT- rich" (rich in adenine and thymine bases), because A-T base pairs have two hydrogen bonds (rather than the three bond in a C-G pair). Once the origin has been recognized, the initiators proteins (origin recognition complex)start the pre-replication complex, which unwind the double- stranded DNA by helicase enzyme. forming
All known DNA replication systems require a free 3' hydroxyl group before synthesis can be initiated. use a primase enzyme to synthesize a short RNA primer(10-20 bp) with a free 3 OH group which is subsequently elongated by a DNA polymerase in this mechanism, In eukaryotes, primase is produced by Pol DNA polymerase helicases, which break the hydrogen bonds holding the two DNA strands together. These two strands serve as the template for the leading and lagging strands, which will be created as DNA polymerase matches complementary nucleotides to the templates; the templates may be properly referred to as the leading strand template and the lagging strand templates. Single strand binding proteins (SSBPs) bind to the single strands of DNA near the replication fork to prevent the ssDNA strands from winding back into a double helix, thus maintaining the strand separation.
2- Elongation step DNA is always synthesized in the 5' to 3' direction. Since the leading and lagging strand templates are oriented in opposite directions at the replication fork, a major issue is how to achieve synthesis of nascent (new) lagging strand DNA, whose direction of synthesis is opposite to the direction of the growing replication fork.
A new DNA strand is always synthesized in a 5 to 3 manner, thus the replication of both the strands goes in two different ways. 1- Leading strand A leading strand is the strand which is run from 5 -3 direction or the direction the same as the replication fork movement. It is synthesized continuously; there are no breaks in-between. This strand is formed as nucleotides are continuously added to the 3 end of the strand after polymerase reads the original DNA template . Only one primer will require here .no Okazaki fragment will formed. 2 DNA polymerase molecules are required for polymerization the two strands the leading and lagging strands are synthesized by Pol (epsilon) and Pol (delta), respectively. Recentlyreplication termination involves a switch from Pol to Pol ; thus, the commonly accepted model is that Pol performs initiation and termination on both strands, as well as the synthesis of Okazaki fragments, whereas Pol elongates only the leading strand. .
1-The leading strand receives one RNA primer while the lagging strand receives several 2- The leading strand is continuously extended from the primer by a high processivity, replicative DNA polymerase, while the lagging strand is extended discontinuously from each primer, forming Okazaki synthesis continues, the original DNA strands continue to unwind on each side of the bubble, forming a replication fork with two prongs. Clamp proteins : it form a sliding clamp around DNA, helping the DNA polymerase maintain contact with its template, thereby assisting with processivity. The inner face of the clamp enables DNA to be threaded through it. Once the polymerase reaches the end of the template or detects double-stranded undergoes a conformational change that releases the DNA polymerase. fragments As DNA DNA, the sliding clamp
2- Lagging strand :it DNA polymerase can extend the RNA primer, adding nucleotides one by one that are complementary to the template strand. ( example: A in the template strand is complement to T in new growing strand, and G in the template strand is complement to C in new growing strand). A DNA polymerase delta ( ) extends the primed segments, forming Okazaki fragments. DNA polymerase will add direction; however, one of the parent strands(lagging) of DNA is 3' to 5' while the other (leading) is 5' to 3'. To solve this problem, replication occurs in opposite directions. lagging strand run away from the replication fork, and synthesized a series of short fragments known as Okazaki fragments, consequently requiring many primers. The RNA primers of Okazaki fragments are degraded by Rnase H. nucleotides in the 5' to 3'
Types of DNA polymerase in Eukaryotic cell The DNA polymerases of eukaryotes are in general less understandable than the DNA polymerases of prokaryotes. Eukaryotic cells have DNA polymerases: DNA polymerase alpha (Pol ), DNA polymerase delta (Pol ) and DNA polymerase epsilon (Pol ), DNA polymerase beta ( poly mitochondria: DNA polymerase Gamma mitochondria also contain their own DNA polymerase that appears to be similar to FIVE polymerases: four major nuclear ), and one found in chloroplasts (Poly ). Plant and 1-Polymerase alpha ( Pol ) : it is the only enzyme has primase activity beside DNA polymerase, it is the only enzyme has primase activity beside DNA polymerase so it is self- primed it will form short primer 12-20 nts called the initiator RNA iRNA 2-Pol Beta processive. polymerase: excision repair and it is not highly active and is not very 3-Pol Gamma polymerase: polymerization the mitochondrial DNA beside repairing by its exonuclease activity 3 5 in mammals. 4-Pol delta and 5- epsilon polymerase :polymerization lagging ( )and leading ( ) strand respectively 5 3 . In eukaryotes.
Termination in Eukaryotic cell Primer removal in eukaryotes is performed by RNase I that removes all the primer leaving only one nucleotide in the junction between 2 nucleotide and the remained one will removed by FenI enzyme. a single nick on the leading strand and several nicks on the lagging strand can be found. Ligase works to fill these nicks in, thus completing the newly replicated DNA molecule. Eukaryote cell initiate DNA replication at multiple points in the chromosome, so replication forks meet and terminate at many points in the chromosome; Because eukaryotes have linear chromosomes, DNA replication is unable to reach the very end of the chromosomes, but ends at the Telomere region of repetitive DNA close to the end. This shortens the telomere of the daughter DNA strand. Shortening of the telomeres is a normal process in Somaticcells. Within the Germ cell line, which passes DNA to the next generation, Telomerase extends the repetitive sequences of the telomere region in the end of the replication the DNA will warp around the basic histones to form the chromatin
The semiconservative replication primer for initiation, and the lagging strand primer to start each Okazaki fragment. Even if an RNA primer can be placed at the very 39 end of the chromosome replicated as the lagging strand, this primer eventually will be lost because no distal Okazaki fragment can be extended through it. The loss of 10 or 12 nucleotides (or more, if the last positioned at the very end) at each cell division should pose major problems for long-lived multicellulareukaryotes. problem is that DNA RNA requires an requires one primer is not
