Translation and

post translational
modification
 Specific learning objective (SLO)
At end of the session the student should be able to …….
o Define translation .
o List the tools required for translation
o Explain the process of protein synthesis
o Explain the post translational modifications
and its significance.
o List the inhibitors of translation.
 Definition of Translation
• It is the process of converting or translating
the information in mRNA into a sequence of
amino acids that makes up proteins.

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 Tools for Protein synthesis
• Ribosomes
• RNAs – mRNA , tRNA rRNA
• Translation factors
• Energy in the form of ATP and GTP
• Aminoacids
Site of protein synthesis -- RIBOSOMES
•The mammalian ribosome has sedimentation coefficients of
80 S unit. It has larger 60 S subunit and another smaller 40S
subunit.
•The bacterial ribosome has sedimentation coefficients of
70 S unit. It has larger 50 S subunit and another smaller
30S subunit.
 mRNA

• Polycistronic mRNA (Prokaryotes) :

MORE THAN ONE POLYPEPTIDE synthesized
from single mRNA.

• Monocistronic mRNA (eukaryotes) :

ONE POLYPEPTIDE synthesized from single
mRNA.
 mRNA
• Nucleotide sequences
present in mRNA concerned
with protein synthesis is
known as GENETIC CODE.

• A series of three adjacent

bases in genetic code forms
a CODON

• Each CODON codes for

single aminoacid.

mRNA
 Genetic Code
• Using the 4 bases A,U,G and C There are 43, or 64 codons are possible.

• AUG acts as the initiator codon.

• 3 Termination codons (UAA , UAG, UGA )

 Genetic Code
AUG codes for methionine in eukaryotes,but
codes for N-formyl methionine in prokaryotes.
 Mitochondrial Genetic code

Genetic code differs slightly from the standard

code
• UGA (Stop codon)is read as Tryptophan
• AGA and AGG (codons for Arginine) read as stop
codons
 Wobble hypothesis
• Why multiple codons can code for a single amino acid ?

• A wobble base pair is a pairing between two nucleotides in RNA molecules that does
not follow Watson-Crick base pair rules.

• Base-pairing in the 3rd position of the anticodon (tRNA )is not strict

• This lets multiple codons be recognized by one tRNA

Base in Bases in 3rd position of

Anticodon Codon
U A or G
G C or U
I (inosine) U, C or A
 tRNA
• It transfer aminoacid to the site of
protein synthesis.

• A triplet of nucleotides in tRNA that

is complementary to the codon in
mRNA—called an anticodon. Amino acid

• Anticodons are present in tRNA

molecule
Anticodon
 tRNA

Pseudo uridine
 Translation factors

The PROTEIN FACTORS required for protein

synthesis are...

• Initiation factors (IF)

• Elongation factors (EF)
• Releasing factors (RF)
 Steps of protein synthesis

• Activation of amino acid.

• Initiation.
• Elongation.
• Termination.
• Post translational modification.
• mRNA translated from 5’ to 3’ end.
• Protein synthesized from N terminal to C
terminal.
 ACTIVATION OF AMINO ACIDS
Enzyme - aminoacyl tRNA synthetase
At least one aminoacyl-tRNA synthetase exist for each
aminoacids.
Recognizes the tRNA and the correct amino acid.
 RECOGNITION OF INITIATION CODON

• The marker sequence for the identification of

INITIATION CODON (AUG )in the mRNA by
RIBOSOMES

• KOZAK CONSENSUS sequence in eukaryotes.

• SHINE-DALGARNO sequence in prokaryotes

 Steps of Initiation
1.Ribosomal Dissociation
2.Formation of 43S pre-initiation complex
3.Formation of the 48S Initiation Complex
4.Formation of the 80S Initiation Complex

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 Ribosomal Dissociation

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 Formation of 43S pre-initiation
complex

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 Formation of the 48S Initiation
Complex

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 Formation of the 80S Initiation
Complex

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 Initiation phase- summary

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 Steps of Elongation
1.Binding of Aminoacyl-tRNA to the A Site
2.Peptide Bond Formation
3.Translocation

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Binding of Aminoacyl-tRNA to the A Site

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 Peptide Bond Formation

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 Translocation

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 Elongation phase- summary

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Termination of
translation
 Termination of
translation-cont...
 POST TRANSLATIONAL MODIFICATIONS

• Proteins synthesised in a few cases may already

be a functionaly active.

• In other cases , proteins synthesised undergo

post translational modification before attaining
biological activity.

• Modifications in proteins occur during their

storage in Golgibodies.
 POST TRANSLATIONAL MODIFICATIONS
• PROTEOLYTIC CLEAVAGE- Proinsulin to insulin.
• HYDROXYLATION- of Pro & Lys as in collagen.
• GLYCOSYLATION- Addition of carbohydrates to
glycoprotein.
• GAMMA CARBOXYLATION- of Glu in clotting factors
with the help of vit-K in prothrombin.
• PHOSPHORYLATION – of certain serine & threonine
residues in casein.
• Formation of S-S bond between two cysteine
residues.
Inhibitors of protein synthesis in prokaryotes.

Tetracyclins Inhibit tRNA binding to

ribosomes.

Chloramphenicol Inhibits the peptidyl

transferase activity of the
50s ribosomal subunit.
Streptomycin Casues misreading of codon

Erythromycin Binds to the 50s subunit &

inhibits translocation.
 Inhibitors of protein synthesis in
Prokaryotes and Eukaryotes

Puromycin It has structural

resemblance to tyrosinyl
tRNA.
Cycloheximide Inhibits peptidyl
transferase in 60s subunit.
Diphtheria toxin Inactivation of EF-2
Thank You