Protein

synthesis- DNA
transcription
and translation
Before that, please recap…RNA and DNA structure
The basic unit in both DNA and RNA

Nucleotide
The polymerization of nucleotides in forming nucleic acids

Backbone of NA=
Phosphodiester bond

Hydrogen
bond

Replace
Thymine in
RNA strand
And that’s the construction of
DNA and RNA
Introduction:

• DNA can control cell activity because it is able to control the production of
various types of protein such as enzyme, collagen, haemoglobin eg.. Which
may effect the metabolisms, defense mechanisms and phenotype of an
organism.

• The information regarding amino acid sequence that will form a protein is
contained within the nucleotide sequence of DNA.

• There are two main processes that a cell needs to use to produce a protein
using the information contained in the DNA sequence that is;
• DNA Transcription
• DNA Translation
The relationship of DNA replication, transcription and translation

Nucleus Cytoplasm

transcription Modified translation

DNA mRNA Protein
mRNA
through
RNA splicing

Jacob and Monod realized that

DNA is in the nucleus, but protein
is synthesize at ribosomes, thus
they conclude mRNA is the one
replication who bring the information to
ribosomes from DNA
DNA transcription
• is a process where RNA is synthesized from information contained in
DNA.

• There are three types of RNA synthesised during the transcription

process:
• Messenger RNA (mRNA) Inside
• Ribosomal RNA (rRNA) nucleus
• Transfer RNA (tRNA)
 Roles of RNA
Types of RNA Function
mRNA -As intermediate in protein synthesis, carries genetic information from
DNA in the nucleus to the ribosomes
-Produce during DNA transcription process

tRNA -Carries amino acids to ribosomes where the mRNA has alligned itself
and functions in recognising codons during protein synthesis
-Involve in DNA translation process

rRNA -The main component of ribosomes

-Involve in DNA translation process
DNA transcription
 Steps
• RNA polymerase bind to DNA promoter site and start to unwind it, and another enzyme unzip it; the
hydrogen bond between bases break.

• Two single stranded DNA formed, only one of them act as the transcribed strand aka template strand
and another one is the non transcribed strand aka non template strand.

• There are free RNA nucleotides in the nucleus; take part in DNA transcription process

• As RNA nucleotides pair up with their complementary one, the sugar phosphate phophodiester bond
is form by the RNA polymerase as it moves in the direction of 3’ to 5’ in the transcribing strand

• The new single stranded molecule formed known as messenger RNA (mRNA) which will leave
through the nucleopore.

• Newly transcribed mRNA is in 5’-3’ direction, will detach from the transcribed DNA template, and will
be modified via RNA processing with RNA splicing before leave the nucleus through nucleopore by
breaking H bond
Modifying the mRNA – RNA processing with RNA splicing
• The original mRNA produced from DNA transcription is known as
primary transcript.

• Modification is needed via RNA processing with RNA splicing

• The introns will be removed and only exons left and join
together to form the modified mRNA for DNA translation

• In some cases, primary transcript can be spliced in different

ways, thus resulting into different mRNA made from the original
primary transcript

• These different mRNA will produce different protein when

translated

• Resulted into one gene can code for variety form from the same
protein
You will expect modified mRNA is shorter
than the transcribed DNA template
The modified mRNA is
synthesised transfer to
cytoplasm through
nucleopore for
translation process
DNA translation
• Before I explain to you the process, you have to know what is the
code in DNA or mRNA represent.
What is genetic code and codon?
• The information of DNA and RNA is carried in the form of codes.

• These “codes” are written into nucleic acid base sequences.

• DNA contain a “triplet genetic code” for every three bases (“codon” in mRNA) code for
one amino acid in a protein.

• There are four different bases in DNA and RNA. Thus, in calculation,the possible
number of genetic code and codon are= 4^3=64 possible codes/codons!!

• Codon refer to the three bases on mRNA while triplet genetic code refer to the three
bases on DNA.
Triplet 1 codon
genetic
code
Some property of the code
• It’s universal- the same code codes for same amino acids in almost all the
organisms.

• The code is degenerate-three bases in DNA codes for one amino acid.

• The code is non-overlapping- example,given a codon with the sequence

AUGUCUAGU in mRNA, it will read as AUG|UCU|AGU but not AUG|UGU|GUC or
AUG|GUG|CUA

• Codon AUG acts as a start signal for synthesis of polypeptides and UAA,UAG and
UGA act as the “stop codon” which signal the termination/completion of
polypeptide synthesis.

• The codon is read (translate) in 5’----3’ direction!!!

Degenerate the codes plsssssssss
DNA triplet codes
RNA codon codes
 rRNA/ ribosomes

60S=Svedberg unit

40S

Total size of rRNA in

eucaryotes 60S+40S= 80S??? Peptidyl transferase
tRNA contain N-base
tRNA Site of amino acid
attachment;
sequences, and fold by correspond with the
pairing through H- mRNA codon, at the
bond and produce a end of 3’ which always
“clover leaf” structure. end with the sequence
CCA need specialized
enzyme to catalyse
the covalent bond
with amino acid

Complementary codon on the

mRNA; at loop 2
 • There are total of three stages in translating the
Stages in mRNA into protein.
• Binding of mRNA to ribosomes.
Translation • Initiation, elongation, and termination of a
polypeptide chain.
Binding of mRNA to ribosomes

• mRNA carries codons complementary to the genetic code on

DNA.
• The 5’ end of the mRNA binds to the smaller ribosomal unit.
• Exposing six bases to large subunit
 • Initiation occurs when the anticodon
UAC of tRNA molecule carrying

Initiation, methionine (binding of amino acid to

tRNA requires ATP and specific
enzyme!) form hydrogen bond to the
elongation and start codon on mRNA on smaller
ribosomal unit.
termination of a • Then, the large ribosomal unit is added
polypeptide to the small subunit to form a
functional ribosome.
chain.
• The initiator codon is now on the P-
site of the ribosome.
Attachment of mRNA on smaller ribosomal unit.
Initiation-when initiator tRNA bind to start
codon AUG at the P-site of
 • A second tRNA with complementary
anticodon enters the A-site and binds
to the second mRNA codon.

Elongation of • The enzyme peptidyl-transferase (on

smaller ribosomal sub-unit)catalyze

polypeptide the formation of the two amino acids

to produce a dipeptide through
peptide bond.
chain
• The covalent bond between amino
acid and tRNA at P-site is broken, the
unbound tRNA in the ribosome is
released back into the cytoplasm.
• The ribosome moves along the mRNA by one codon(3 bases).The second tRNA
now is located at the P-site of the ribosome leaving the A-site vacant for
incoming tRNA bringing the amino acid.

• A third tRNA with anticodon complementary to the new codon enters and
occupies the A-site. A peptide bond is formed to link the second amino acid
with the third amino acid to form a tripeptide . The second uncharged tRNA is
then released back to the nucleoplasm
• The process of the translation is repeated.

• Each time a new aminoacyl-tRNA with complementary anticodon

with the codon on mRNA on the ribosome enters will form a peptide
bond in between adjacent amino acids.

• As process continues, a polypeptide chain is formed.

Termination of polypeptide chain
• When the ribosome encounters a terminator (stop codon-UAA,UAG
or UGA), a protein called the releasing factor binds to the stop codon.

• Hydrolysis of the bond between the tRNA and the last amino acid in
the polypeptide chain occurs.

• The complete polypeptide chain is released from the ribosome

• The ribosome subunit are disintegrate and released and reused again.
Termination of polypeptide chain
Polyribosome.
• Many ribosomes forming a polyribosomes are usually found on mRNA
molecule.

• It allows several polypeptide chains to be assembled from one mRNA

molecule within a short time.

Not really in syllabus anymore

Not really in syllabus
anymore
Gene mutation
• Mutation
• Chromosome mutation - change in structure/ number of chromosomes

• Gene mutation – change in the base/nucleotide sequence in DNA

• Base substitution
• Base deletion
• Base insertion (aka addition)
Base substitution
• Substitute a base on a DNA with another base

• Eg :

• Change a DNA triplet code, change a codon on mRNA, change a tRNA bringing a
different amino acid during translation, change sequence of amino acid (primary
structure)

• Some causes “silent mutation” as different triplet code might code for same
amino acid eg :
Case study : Sickle cell anaemia
• B-polypeptide affected via base substitution
Base insertion – causes
frameshift mutation

• Frame shift mutation start from the point

of base addition.
• Code unchanged before point of mutation
• Code changes start at the point of
mutation
• The sequence of amino acid start from the
mutation point will most likely be
different; same before point of mutation
• May results same length of polypeptide or
shorter polypeptide chain produced ;early
stop code appear
DNA triplet codes RNA codon codes
Base deletion – causes frameshift mutation
• Frame shift mutation start from the point of base addition.
• Code unchanged before point of mutation
• Code changes start from the point of mutation
• The sequence of amino acid start from the mutation point will most likely
be different; same before point of mutation
• Sequence of polypeptide at least shorten by 1 amino acid (lack of a base;
one code will disappear) ;or shorter polypeptide due to early stop code
appear
DNA triplet codes RNA codon codes
DNA being stable
• sequence will not (spontaneously) change / AW ; A decreases chance
of mutation
• (so) gene products / proteins, produced will always be functional ;
• maintains all, genetic information / AW, throughout life of cell ;
• same, genetic information / AW, passed on to, daughter cells /
offspring ;
• AVP ; e.g. maintains size so still enclosed within nucleus
Importance of H bond in DNA?
• hydrogen bonds hold (two), polynucleotides / strands / chains, together ;

• hold, (complementary) nucleotides / base pairs, together

A ora e.g. prevents, unwinding / strand separation

• (many hydrogen bonds) give stability / DNA is stable molecule / DNA is long lasting /
AW ; ignore ref. to strength

• can be broken for, transcription / replication ;

• ref. to (double) helix ;

mRNA being labile
• translation / protein synthesis, will stop when mRNA breaks down ;
• allows re-use of nucleotides (for other mRNA) ;
• ref. to control of gene expression ; A prevents too much product
forming
• ref. to control of cell activity / fast response to changing rquirements ;
• ref. to efficiency in energy use ;
Role of mRNA?
• 1 translation ; R if transcription given as well, unless in correct context
A use of, nucleotide / base, sequence, to make, amino acid chain / polypeptide / protein
I protein / polypeptide, synthesis

• 2 moves towards / combines with, ribosome ;

• 3 ref to small and/or large sub-units ; I small / large ribosome

• 4 codon(s) ; only accept in correct context

• 5 transfer / t, RNA, bringing, amino acid(s), to mRNA / ribosome

• 6 anticodon(s) ; only accept in correct context

• 7 (complementary) base pairing ;

• 8 any e.g. of codon:anticodon base pairing ; need six bases

• 9 ref to polyribosome(s) / used by many ribosomes ;

• 10 (mRNA short-lived) ref to production of protein for short period of time ;

Role of ribosomes in DNA translation/protein synthesis

1 translation (in correct context) / genetic code used to make a sequence

of amino acids /AW ;
2 attach / assemble around / moves along / AW, mRNA ;
A ref. to bind mRNA / mRNA ‘lies within’ the ribosome
3 provides two sites for tRNAs carrying amino acids to bind to mRNA
4 binding / pairing / AW, between anticodon on tRNA to codon on
mRNA ;
5 (catalyse) formation of peptide bond (to form polypeptide) ;
6 any further detail of translation ;
e.g. peptidyl transferase
ribosome moves along one codon at a time
start codon is AUG
stop codon in context
The role of tRNA in DNA translation/ protein
synthesis
• tRNA combines with specific amino acid / carries
specific amino acid /activated
• tRNA to ribosome;
• idea of, adjacent / two, amino acids AND codon-
anticodon binding ;
• peptide bond formation / ref. elongation, (to form
polypeptide) ;
• A ref to roles in two sites on ribosome at P and A site
(and E)
• leave ribosome after amino acid joins
polypeptide(continually reused);

• Do S13_qp_23 question 2