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

• •

• •

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

tRNACommon Features

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

mRNA

5’

3’

A

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

G C G A

U A

Met

Gly

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

3’

Met

Gly

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

3’

Gly

Cys

5’
mRNA C C UA C A

3’
A U G G G A U G U A A G C G A

Gly

Cys

mRNA

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

3’

Cys

Lys

Lengthening polypeptide (amino acid chain)

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

3’

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

mRNA

5’
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

mRNA

5’
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 peptidyltRNA 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 Nterminal 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 Show

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