Topic to be discussed today:

1. Wobble hypothesis

2. General transcription process (summary)

3. Inhibitors of transcription and translation

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Wobble hypothesis
Suggested by Watson and Crick.

This explains the redundancy of genetic code.

By model building, he was able to suggest that the 3rd base
of the 5’- anticodon is able to go undergo more movement
compared to the other two bases.
This above model would lead to the formation of non-
standard base pairs.

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Wobble hypothesis
Non-standard base pairing occurs as long as the distance
between the ribose units were close to normal.

No purine-purine or pyrimidine-pyrimidine base pairs are

allowed due to incorrect ribose distance.

No single tRNA could recognize more than one three codons.

So, atleast 32 tRNA’s would be needed to decode the 61

codons (excluding stop codon).

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Wobble hypothesis
tRNA’s are able to recognize either one, two or three codons
depending on their wobble base (the 5’-anticodon base).

5' anticodon base predicted 3' codon base read observations

A U A converted to I by anticodon deaminase

C G no wobble, normal base pairing

G C&U G and modified G can base pair with C and U

U A&G U not found as 5' anticodon base

I C&U wobble as predicted, Inosine (I) can recognize 3' -A, -C or -U

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Wobble hypothesis
The wobble nucleoside will never be A as it is converted to
Inosine which can then pair with A, C or U.

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 General Translation
mechanism

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overview
Protein synthesis occurs in Three steps:
Initiation: the assembly of ribosomes on an mRNA
molecule.

Elongation: repeated cycles of amino acid delivery, peptide

bond formation and movement along the mRNA
(translocation).

Termination: release of the new protein chain.

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Initiation
Prokaryotes need: large and small ribosome subunits, mRNA, the
initiator tRNA, three Initiation Factors (IF’s) & GTP.
IF1 and IF2 bind to 30s subunit and prevent large subunit binding.
IF2 + GTP bind and help the initiator tRNA to bind later.
this small subunit complex can now attach to an mRNA via a
ribsome-binding site.
The tRNA forms base pairs with the AUG initiator codon which
releases IF3 , creating 30s initiator complex.
Large subunit binds and displaces IF1 & IF2 + GDP, giving 70s
initiation complex (fully assembled ribosome at correct position
of the mRNA).

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Elongation
It involves three factors called Elongation Factors (EF’s),
EF-Tu, EF-Ts and EF-G, GTP, charged tRNA’s and the 70s
initiation complex (or its equivalent).

It takes place in three steps:

A charged tRNA is delivered as a complex with EF-Tu and

GTP. GTP hydrolyzed and EF-Tu.GDP is released, this is re-
used with the help of EF-T’s and GTP (via the EF-Tu-EF-Ts
exchange cycle).

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Elongation
Peptidyl transferase makes the peptide bond by joining the
two adjacent amino acids without input of more energy.

Translocase (EF-G), with energy from GTP, moves the

ribosome one codon along the mRNA, ejecting the
uncharged tRNA and transferring the growing peptide
chain to the P-site (peptidyl site, it also has aminoacyl site
or A-site).

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Termination
Release factors (RF1 & RF2) recognize the stop codons and
helped by RF3 make peptidyl transferase join the
polypeptide chain to a water molecule, thus releasing it.

Ribsomes release factor helps to dissociate the ribosome

subunits from the mRNA.

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Inhibitors of transcription and translation

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Inhibitors of transcription and translation
Many antibacterial agents have been isolated from fungi
(antibiotics).
These inhibit protein synthesis.
Examples are as follows:
1. Streptomycin and neomycin – bind to a particular protein in the
30s particle and prevent binding of the tRNAfmet to the P- site.
2. Tetracyclines inhibit binding of charged tRNA.
3. Lincomycin and chloramphenicol inhibit the peptidyl transferase.
4. Puromycin – premature chain termination.
5. Erythromycin- binds to a free 50s particle and prevents the
formation of 70s ribosomes.

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Inhibitors of transcription and translation
Most of the inhibitors for prokaryotes do not affect
eukaryotes and then only they will be of some clinical use
(ribosome binding).

Incase of Diphtheria: bacteria exerts a pathogenic effect by

excreting inhibitors for mammalian protein synthesis. In
this case it binds to the factors necessary for movement of
mammalian ribosomes along the mRNA.

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 The End

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