Name: Mirza Omer

Roll no: 29

Assignment no :5 (DNA Replication – DNA

transcription)

Course: Molecular Genetics

Date :4/20/2020

Submitted to : Dr Shaukat Malik

Department of Biotechnology capital university of science

and technology
N Characteristics Replication
o
1 DNA replication is the process of
making two daughter strand
Definition where each daughter strand
contains half of the original DNA
double helix.
2 To conserve the entire genome
Purpose
for next generation.
3 Occurs in the S phase of cell
Occurrence
cycle.
4 Enzymes Required  Helicase (unwinds the DNA
double helix)
 Gyrase (relieves the
buildup of torque during
unwinding)
 Primase (lays down RNA
primers)
Transcription
Transcription is the process of
synthesis of RNA using DNA as a
template.
To make RNA copies of
individual genes.
Occurs in the G1 and G2 phases
of cell cycle.
Enzymes along with transcription
factors are need to initiate
eukaryotic transcription
TFIID,TBP,TFIIA,TFIIA,TFIIE,TFIIF.
Enzymes involved in prokaryotic
DNA Transcription is RNA
polymerase .
  DNA polymerase III (main
DNA synthesis enzyme)
 DNA polymerase I
(replaces RNA primers with
DNA)
 Ligase (fills in the gap
5 Occurs in the S phase of Occurs in the G1 and G2 phases
Occurrence
cell cycle. of cell cycle.
6 dATP, dGTP, dTTP and
ATP, UTP, GTP and CTP serve as
Raw Materials dCTP serve as raw
raw materials.
materials.
7 Occurs along the strands
Occurrence Occurs along one strand of DNA.
of DNA.
8 Occurs in preparation for Occurs in preparation for protein
Occurrence
cell division. translation.
9
Bond Replicated DNA strand remains Transcribed RNA strand
hydrogen bonded to its template separates from its DNA template
DNA strand. strand.

10
Primers It require RNA primer to start
No primer is required to start.
replication.

11 Main Enzyme

DNA polymerase RNA polymerase

One of the key molecules in DNA
replication is the enzyme DNA
polymerase. DNA polymerases are
responsible for synthesizing DNA: they
add nucleotides one by one to the
The main enzyme involved in
transcription is RNA polymerase,
which uses a single-stranded DNA
template to synthesize a complementary
strand of RNA. Specifically, RNA
 growing DNA chain, incorporating only
those that are complementary to the
template.
Starting DNA Replication always starts at specific
12
replication locations on the DNA, which are
called origins of replication and are
recognized by their sequence.
The Replicon Model
All the DNA replicated from a particular
origin as a replicon.
For example, the single chromosome
found in E. coli cells it has only one
origin of replication, the entire
chromosome is a single replicon.
In contrast, multiple origins of
replication divides each eukaryotic
chromosome into multiple replicons—
one for each origin of replication.
The replicon model proposed two
components that controlled the initiation
of replication: the replicator and the
initiator
The replicator is defined as the entire
set of cis-acting DNA sequences that is
sufficient to direct the initiation of DNA
replication.
The second component of the replicon
model is the initiator protein.
This protein specifically recognizes a
polymerase builds an RNA strand in the
5' to 3' direction, adding each new
nucleotide to the 3' end of the strand.
Replication always starts at specific
locations on the DNA, which are called
Promoter Sequences / Recognition
sites
Each gene (or group of co-transcribed
genes, in bacteria) has its own promoter.
Once bound, RNA polymerase separates
the DNA strands, providing the single-
stranded template needed for
transcription.
Promoters in bacteria
A typical bacterial promoter contains
two important DNA sequences, the -10
and -35 elements.
Promoters in humans
In eukaryotes helper proteins called
basal (general) transcription factors bind
to the promoter first, helping the RNA
polymerase in your cells get a foothold
on the DNA.
Many eukaryotic promoters have a
sequence called a TATA box. The
TATA box plays a role much like that of
the -10 element in bacteria. It's
recognized by one of the general
transcription factors, allowing other
 DNA element in the replicator and transcription factors and eventually
activates the initiation of replication. RNA polymerase to bind
Initiator proteins have been identified in
many different organisms, including
bacteria, viruses, and eukaryotic cells,
they all select the sites that will become
origins of replication.
The single replicator required for E. coli
chromosomal replication is called oriC.
There are two repeated motifs that are
critical for oriC function (Figure 8-24a).

The 9-mer motif is the binding site for

the E. coli initiator, DnaA, and is
repeated five times at oriC.
The 13-mer motif, repeated three times,
is the initial site of ssDNA formation
during initiation.

13 Opening structure
Of DNA Replication Bubble
As the DNA opens, two Y-shaped
structures called replication forks are
formed, together making up what's called transcription stage of protein synthesis
a replication bubble. The replication
forks will move in opposite directions as
replication proceeds.
Elongation During elongation, a primer sequence is
14
added with complementary RNA
nucleotides, which are then replaced by
Transcriptional
Bubble
The Open complex forms during the
after the less stable closed complex. ...
The change from a closed complex to an
open complex involves the unwinding of
the DNA strands (forming a
transcription bubble).
During elongation, RNA polymerase
"walks" along one strand of DNA,
known as the template strand, in the 3' to
 DNA nucleotides. During elongation the
leading strand is made continuously,
while the lagging strand is made in
pieces called Okazaki fragments
Termination The ends of the parent strands consist of
15
repeated DNA sequences called
telomeres. ... Once completed, the parent
strand and its complementary DNA
strand coils into the familiar double helix
shape. In the end, replication produces
two DNA molecules, each with one
strand from the parent molecule and one
new strand.
5' direction. For each nucleotide in the
template, RNA polymerase adds a
matching (complementary) RNA
nucleotide to the 3' end of the RNA
strand.
RNA polymerase will keep transcribing
until it gets signals to stop. The process
of ending transcription is called
termination, and it happens once the
polymerase transcribes a sequence of
DNA known as a terminator.
Termination in bacteria
There are two major termination
strategies found in bacteria: Rho-
dependent and Rho-independent.
In Rho-dependent termination, the
RNA contains a binding site for a
protein called Rho factor. Rho factor
binds to this sequence and starts
"climbing" up the transcript towards
RNA polymerase.
Rho-dependent termination. The
terminator is a region of DNA that
includes the sequence that codes for the
Rho binding site in the mRNA, as well
as the actual transcription stop point
(which is a sequence that causes the
RNA polymerase to pause so that Rho
can catch up to it). Rho binds to the Rho
binding site in the mRNA and climbs up
the RNA transcript, in the 5' to 3'
direction, towards the transcription
bubble where the polymerase is. When it
catches up to the polymerase, it will
cause the transcript to be released,
ending transcription.
Rho-dependent termination. The
terminator is a region of DNA that
includes the sequence that codes for the
Rho binding site in the mRNA, as well
as the actual transcription stop point
(which is a sequence that causes the
RNA polymerase to pause so that Rho
can catch up to it). Rho binds to the Rho
binding site in the mRNA and climbs up
the RNA transcript, in the 5' to 3'
direction, towards the transcription
bubble where the polymerase is. When it
catches up to the polymerase, it will
cause the transcript to be released,
ending transcription.
When it catches up with the polymerase
at the transcription bubble, Rho pulls the
RNA transcript and the template DNA
strand apart, releasing the RNA
molecule and ending transcription.
Another sequence found later in the
DNA, called the transcription stop point,
causes RNA polymerase to pause and
thus helps Rho catch up.^4
4
start superscript, 4, end superscript

16 Proofreading Present
Ability Proofreading (biology) ... In bacteria, all
three DNA polymerases (I, II and III)
have the ability to proofread, using 3' →
5' exonuclease activity. When an
incorrect base pair is recognized, DNA
polymerase reverses its direction by one
base pair of DNA and excises the
mismatched base.
Rho-independent termination depends
on specific sequences in the DNA
template strand. As the RNA
polymerase approaches the end of the
gene being transcribed, it hits a region
rich in C and G nucleotides. The RNA
transcribed from this region folds back
on itself, and the complementary C and
G nucleotides bind together. The result
is a stable hairpin that causes the
polymerase to stall.
Absent
All nucleic acid polymerases insert
incorrect nucleotides during chain
elongation. ... This high rate of mutation
comes from the lack of proofreading
ability in RNA polymerases. These
enzymes make mistakes, but they can't
correct them. Therefore the mutations
remain in the newly synthesized RNA.

17 mRNA, tRNA, rRNA

Products Two Daughter Strands and non-coding RNA(
like microRNA)
18 Greater part of the
product passes from
Products Products remain within nucleus.
nucleus into the
cytoplasm.
 19 Products are
Products Products are not degraded. degraded after their
function of over.
20 It involves copying of
It involves copying of the entire
copying certain individual
genome.
genes only.
21 It involves unwinding
Unwinding It involves unwinding and
and splitting of only
and splitting of the entire DNA
those genes which
Splitting molecule.
are to be transcribed.
22 It produces primary
It produces normal DNA RNA transcript
processing molecules that do not need any molecule which needs
processing. processing to acquire
final form and size

References:
https://www.khanacademy.org/science/biology/dna-as-the-genetic-material/dna-
replication/a/molecular-mechanism-of-dna-replication

https://www.khanacademy.org/science/biology/gene-expression-central-dogma/transcription-of-
dna-into-rna/a/overview-of-transcription