DNA

Translation
Translation
Involves the following components:
• Ribosomes( 4 types RNA + 82
proteins)
- Large subunit - small subunit
• Accessory protein factors
1- initiation 2- elongation 3- termination
• Activated amino acids
- set of tRNA
- set of aminoacyl-tRNA synthetase
- set of aminoacyl-tRNA
Ribosomes
Made up of rRNA & proteins
 Prokaryote ( 70S ribosome )

 Small subunit 30S

 Large subunit 50S

 Eukaryotes ( 80S ribosome )

 Small subunit 40S
 Large subunit 60S
 Genetic
Code
 Triplet code (three bases).
 64 ways to code for 20 amino acids.
 All proteins start with methionine
(AUG).
 All proteins end with one of three
stop codons ( UAA, UAG , UGA ).
 The code is said to be degenerate –
the last base in the coding triplet
often does not affect the amino acid
to be added.
The Genetic Code
Genetic code
 tRNACommon
Features
• a 5’ terminal phosphate group
• a 7 bp stem where amino acid
binds
• a 3 or 4 bp stem ending in a loop
• a 5 bp stem ending in a loop that
contains the anticodon, the triplet
that is complementary to the
codon
• all tRNA terminate in the sequence
 Polypeptide
Synthesis
• Proceeds from N-terminus to
C-terminus
• Ribosomes read mRNA in the
5’ - 3’ direction
• Active translation occurs on
polyribosomes
• Chain elongation occurs by
the linkage of the growing
polypeptide to the incoming
tRNA’s amino acid residue
TRANSLATION ELONGATION

5’ 3’
mRNA

A U G G G A U G U A A G C G A

U A C C C U

Met Gly
 5’ 3’
mRNA

A U G G G A U G U A A G C G A
U A C C C U

Met Gly
 5’ 3’
mRNA
A U G G G A U G U A A G C G A
C C U A C A

Gly Cys
 5’ 3’
mRNA
A U G G G A U G U A A G C G A

C C UA C A

Gly Cys
 5’ 3’
mRNA
A U G G G A U G U A A G C G A

A C A U U C

Cys Lys

Lengthening
polypeptide
(amino acid chain)
 5’ 3’
mRNA
A U G G G A U G U A A G C G A

A C A U U C

Cys Lys
 5’
mRNA
A U G G G A U G U A A G C G A U A A

U U C

Lys
 Stop codon

5’
mRNA
A U G G G A U G U A A G C G A U A A

U U C

Lys

Release factor
 Stop codon

5’
mRNA
A U G G G A U G U A A G C G A U A A

Release
factor
Once stop codon is reached,
elements disassemble

Release
factor
Protein Synthesis: Elongation
((1
Peptidyl transferase is not your typical
enzyme. Peptidyl transferase is the
enzyme responsible for peptide bond
formation during protein synthesis. This
enzyme catalyzes the reaction between
the amino group of the aminoacyl-tRNA,
forming a peptide bond from an ester
bond. The enzyme activity is located in
the ribosome, but none of the ribosomal
proteins has the capacity to catalyze this
reaction.
Protein Synthesis: Elongation
((2
In fact, when ribosomes are stripped
of all of their associated proteins,
the rRNA appears to remain capable
of converting an ester bond to a
peptide bond. These observations
have led investigators to
hypothesize that peptidyl
transferase activity is contained in
the rRNA, rather than in the proteins
that associate with ribosomes.
Protein Energy Cost

 There is a considerable use of energy

in protein synthesis
 Four ATPs are used in each
polypeptide bond formation
 In addition post translational
modification adds to the expenditure
of energy
Protein Synthesis in
Mitochondria
 Independent protein synthesis
apparatus
 Most steps in mitochondria are
similar to that in the cytosol
 Ribosomes are smaller & the mRNA
is shorter
 N-formyl methionine is coded by AUG
 Most mitochondrial proteins are
encoded by the nuclear DNA
Antibiotics
 Some antibiotics are selective in their
ability to inhibit protein synthesis in
prokaryotes than eukaryotes
 These antibiotics are extremely
useful in clinical practice
Antibiotics action at Small
subunit 1
 Streptomycin:(tuberculosis) binds the
Streptomycin:(tuberculosis) binds the
small subunit of 70S ribosome & thus
interfering with initiation of protein
synthesis & causes misreading of mRNA.
Mutation in the rRNA can confer resistance
to streptomycin
 Neomycin : (skin ) causes mistranslation
by interacting with the small ribosomal
subunit at different site from above
 Kasugamycin :(antifungal ) binds to small
subunit and inhibits the initiation of
translation
Other antibiotics action sites
 Tetracyclins : (acne & bronchitis) binds
directly to ribosome & interfer in
aminoacyl-tRNA binding
 Puromycin : binds at ribosomal A-site &
acts as an accepter in peptidyl transferase
reaction & thus terminate translation
prematurally
 Chloramphenicol : (typhoid & enteric )
binds peptidyltransferase & thus peptidyl-
tRNA remains bound to ribosome
 Erythromycin : ( chest infection ) interfers
with translocation on prokaryotic ribosome
Toxins
 Diphtheria toxins : inhibits
translocation in eukaryotic ribosome
 Ricin : (caster bean) cleaves a single
adenine from large subunit of
ribosome & cause its inactivation
 α-sarcin : (fungal RNases ) cleaves
large subunit rRNA at a single site &
inactivate it
Protein Maturation 1
 Some proteins emerge from the
ribosome ready to function, while
others undergo a variety of post
transcriptional modifications.
 Signal polypeptide near the N-
terminal for secretory proteins.
 Glycosylation of proteins.
Protein Maturation 2
 Partial hydrolysis of proteins as the
case in insulin.
 Modification of amino acids such as
methylation, acetylation and
hydroxylation such as in collagen
End Of
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