CHAPTER 4A

DNA Replication & Protein Synthesis

REPLICATION PROCESS
• Before the cell divides (mitosis and
meiosis), the DNA must be replicated.
• Process of DNA replication in the
nucleus during interphase (S phase)
• DNA enzymes break the bonds between two nucleotides
of a DNA molecule The two strands (parent
strands) unwind from each other, exposing their
nucleotides.
• The both parent strands act as a template for formation of
the new strands Base pairing rule applies.
• For each nucleotide of the parent strands, a
complementary base is delivered to them.
• As the nucleotides form bonds, the next nucleotide is
delivered until the whole strands are replicated.
DNA REPLICATION
DNA replication requires:
1. Double helix DNA template
2. Nucleotides (deoxy nucleoside triphosphate)
 dATP, dCTP, dGTP, and dTTP.
 2 extra phosphate group broken off to release energy
used to form a sugar-phosphate bond, joining the new
nucleotide to the growing polynucleotide chain.
3. Enzymes
i. Helicase
• Unwinds & unzips (by breaking H-bonds) DNA
double helix to 2 straight chains/strands.
 5’
DNA REPLICATION

3. Enzymes (cont…..)
3’
ii. DNA Polymerase
• Always add a phosphate group on C5’ of the dNTP to the C3’ OH
group of the terminal nucleotide of the growing DNA chain.
• Therefore, the complementary DNA chain grows from 5’ to 3’
direction, starting from 3’ end of the template.
• DNA polymerase must detect the presence of hydroxyl group
on C3’ before they can add new nucleotides.
iii. DNA Ligase
• Join newly formed DNA fragments on the lagging strand from 3’
end to 5’ end.
REPLICATION
PROCESS
As soon as the strands are
replicated, they twist into a
double helix.
The result is half new (Daughter
strands) and half old strands
(Parent strands).
This replication is known as
“Semiconservative
Replication”.
Following replication, each new
DNA molecule consists of one
strand from the original duplex
and one newly constructed
strand.
NUCLEOTIDE ADDITION
• Each DNA strand has a 3’ end and a 5’ end.
• The 3’ end refers to the 3rd carbon of the sugar molecule
which is attached to a hydroxyl group (OH).
• The 5’ end refers to the 5th carbon of the sugar molecule
which is attached to a phosphate group.
• Nucleotides are added to the 3’ end of the growing DNA
strands.
• So, one daughter strand is synthesized continuously and
the other is in short strands (Okazaki fragments).
• DNA polymerase activates the attachment of nucleotides
to the parent strands.
• DNA ligase fills the gap between the short strands into a
continuous strand.
Mechanism of replication
DNA consists of two strands
that run in opposite directions
(antiparallel) .
DNA polymerase molecules are
only capable of moving along a
template in one direction, toward
the 5’ end of the template.
The two newly assembled strands
(daughter strands) grow in
opposite directions, one growing
toward the replication fork, and
the other growing away from it.
One strand is assembled in
continuous fashion, the other in
segments that must be joined
together by an enzyme.
DNA REPAIR
• DNA polymerase, DNA ligase and other enzymes
also engage in a process called DNA repair.
• If the sequence of bases in one strand of a double
helix becomes altered, DNA polymerase can “read”
the complementary sequence on the other strand.
• With the aid of other repair enzymes, the original
base sequence can be restored.
PROTEIN SYNTHESIS
 Learning Outcomes
• To understand genetic coding in relation with
complementary base pairing.

• To understand the replication, transcription and

translation concepts and processes.

• To explain the relationship among replication,

transcription and translation processes in living
cell.
 CONCEPT

‘DNA makes RNA

and
RNA makes protein’

Polypeptide
synthesis
Polypeptide
GENE CONCEPT
One DNA molecule contains genetic information in the form of
many genes.
Each gene has a specific sequence of bases which codes for
one polypeptide.
The polypeptide formed will become a functional protein, such
as an enzyme.
Through the protein or enzyme, a biochemical is produced.
The gene concept can be represented as follow:
The relationship of gene and
protein.
 GENETIC CODE
• Genetic code is a form of three bases in the DNA that
represents one amino acid in a polypeptide.
• The genetic codes found in mRNA are complementary to
those of a DNA reference strand.
• The characteristics of a genetic code:
(a) It is a triplet code – three bases in the DNA
template codes for an amino acid.
Eg TTC coded for Phe

(b) It is degenerative – more than one code codes for

one amino acid.
Eg TTC and TTT coded for Phe
GENETIC CODE
(c) It is non-overlapping – the base in the code is read
only once.
Eg TTCTTA coded for Phe Leu, NOT Phe(TTC) Ser(TCT)

(e) It is comaless – no base acts as a border between two

codes.
Eg TTCTTAC coded for Phe(TTC) Ser(TTA),NOT
Phe(TTC) (with T in the middle acts as a coma) TAC(Thy)

(f) There are three ‘nonsense’ codes that act as ‘full

stop’.
Eg TAA, TAG, TGA is STOP codon
CONT… GENETIC CODE
PROTEIN SYNTHESIS
• Divided into two steps: transcription and
translation.
• Requires three types of RNA: messenger RNA
(mRNA), ribosomal RNA (rRNA) and transfer RNA
(tRNA).
RNA Molecules
 Each RNA molecule has a 5C sugar (ribose), a
phosphate group and a nitrogenous base.
 There are four types of bases: adenine, guanine,
cytosine and uracil.
 Base pairing: adenine with uracil and guanine with
cytosine.
Structure of one of the four nucleotides of
RNA.
The other three have a different base (adenine,
guanine, or cytosine instead of the uracil shown
here).
Overview of Protein Synthesis

Overview of
transcription
and
translation.
These two
steps leads to
protein
synthesis as
they occur in
eukaryotic
cells.
 TRANSCRIPTION
 The mechanism by which the base sequence of a gene is
converted into the complementary base sequence of mRNA .
(synthesis of mRNA)
 The DNA double helix unwinds & unzips (breakage of H-bonds)
by RNA polymerase at the beginning of a gene.
 RNA Polymerase binds to the template strand (3’ END) & using it
as a template, corresponding free RNA nucleotides triphosphate
are added one by one continuously from the beginning till the
end of the gene in a 5’ 3’ direction
 RNA Polymerase then disengage from the DNA & transcript is
released
(1◦ RNA transcript is processed to form mRNA, rRNA, or tRNA
 These RNAs leave the nucleus into the cytoplasm through the
nuclear pores.
 The DNA strands ‘zip up’ & wind up again into DNA double helix
TRANSCRIPTION
Transcription: the synthesis of an RNA molecule on a DNA template.
 STRUCTURE OF MRNA
• Newly formed mRNA transcript is an unfinished
molecule that must be modified.
• Enzymes attach a cap (5’cap of 7-
methylguanosine) to the start of the pre-mRNA
and a tail (poly-A tail) to the end of it.
• Exons (protein coding region) and introns
(non-coding region) which must be removed
along will snipped out introns and the exons will
be joined together. This will resulted in correct
folding of protein after translation.This process is
called RNA splicing must take place before
mRNA leaves the nucleus in mature form.
 Gene splicing

Introns Introns

Transcription and modification of newly formed mRNA in the nucleus.

The genetic code
(as written in
mRNA).
 STRUCTURE OF tRNA
• They are free molecules in the cytoplasm.
• One end of the molecule is a structure called the “hook”, where it attaches
an amino acid.
• The other end has three nucleotide bases called anticodons.
• Anticodons correspond to the codons of the mRNA.
• The anticodon of the tRNA will determines the right amino acid joins
to a specific tRNA.
• Each amino acids is attached to its specific tRNA by a specific
aminoacyl-tRNA synthetase
• This produces an amino acid-tRNA complex known as aminoacyl-tRNA
Model of tRNA.
(a) Structural features common to all
tRNAs.

(b) (b) Simplified model of tRNA that is used

(a) in illustration. The “hook” at one end is
the site to which a specific amino acid
can become attached.
STRUCTURE OF rRNA (RIBOSOME)
• Ribosomes are made of ribosomal RNA and proteins.
• There are two subunits of ribosome.
• Small subunits (40S) and large subunits (60S).
• Ribosomes are the sites for protein synthesis.
• Here, the mRNA ‘code’ is translated into a sequence of
amino acids in a polypeptide chain.
• The mRNA binds to the small subunit. Six bases, at a
times, are exposed to the large subunit.
• The tRNA will deliver the amino acids to the ribosomes one
by one.
TRANSLATION
Stages of
translation, the
second step of
protein synthesis:

(a) Initiation stage.

TRANSLATION
 The mechanism by which the sequence bases in a mRNA is
converted to a sequence of a.a. to form a polypeptide chain
(synthesis of polypeptide).
 Ribosome (1small+1large subunit) bind to the start codon (AUG)
on mRNA. 2 codons of mRNA are exposed in ribosomes.
 met-tRNA (methionine-carrying tRNA) binds to the 1st triplet
codon by its complementary anticodon.
 The 2nd triplet codon on mRNA is exposed for another incoming
aminoacyl-tRNA showing the complementary anticodon to bind
to it.
 When the 2nd aminoacyl-tRNA sits into the ribosomes a peptide
bond is formed btw the two a.a., catalysed by an enzyme
(peptidyl transferase).
 The 1st tRNA leaves mRNA to pick up another specific a.a.
TRANSLATION
 The ribosome moves along the mRNA for a distance of 1 triplet
codon & exposed the next triplet codon for the appropriate
tRNA to bind in.
 Specific sequence of a.a. is assembled as the ribosome moves
along the mRNA.
 Many polypeptide chains can be made at the same time as
many ribosomes move along the mRNA. (seen as
polyribosomes)
 When the ribosomes encounters the stop codon, there is no a.a
to bind to it.
 The completed polypeptide is release from the ribosome & the
mRNA.
CONT… TRANSLATION

(b) Chain
elongation
stage.
CONT… TRANSLATION
(b) Cont… Chain elongation stage.
CONT… TRANSLATION

(c) Chain termination

stage.