Genetic Code and Translation
GENETIC CODE
The information needed to direct the synthesis of protein is
contained in the mRNA in the form of a genetic code.
The genetic code is the system of nucleotide sequences of
mRNA that determines the sequence of amino acids in
protein.
Codons are a group of three adjacent bases that specify the
amino acids of protein.
Dictionary of amino acid code words in mRNAs.
Characteristics of Genetic Code
• Universal
• Degenerate
• Unambiguous
• Nonoverlapping and commaless.
The code is degenerate but unambiguous:
As there are 61 codons for 20 amino acids, one
amino acid has more than one codon and the code
is referred to as degenerate.
Although an amino acid may have more than one
codon, each codon specifies only one amino acid.
Thus, the genetic code is unambiguous.
The code is almost universal
Exceptions to the universality of the genetic code
are found in human mitochondria
• UGA codes for tryptophan instead of stop codon
• AUA codes for methionine instead of isoleucine
• CUA codes for threonine instead of leucine.
Code is non-overlapping and without punctuation
During translation, code is read sequentially,
without spacer bases, taken 3 at a time, e.g.
A U G C U A G A C U U U is read as:
AUG / CUA / GAC / UUU
Wobble Hypothesis for Codon-anticodon Interactions
The third base of most codons pairs rather loosely with
the corresponding base of its anticodon, the third base of
such codons (and the first base of their corresponding
anticodons) “wobbles”.
Crick proposed an assumption to explain these data on
the basis of set of four relationships called the “wobble
hypothesis”.
Pairing relationship of codon and anticodon.
Three different codon paring of arginine is possible
when the tRNA anticodon contains inosine (I).
TRANSLATION
(PROTEIN BIOSYNTHESIS)
The pathway of protein Biosynthesis is called
translation.
Translation is the process by which ribosomes
convert the information carried by mRNA in
the form of genetic code to the synthesis of
new protein.
Translation occurs in cytosol on ribosomes and
is guided by mRNA.
Basic plan of protein synthesis in eukaryotes is
similar to that in prokaryotes.
Basic Requirements for the Translation
mRNA
tRNAs
Ribosomes
Energy in the form of ATP and GTP
Enzymes and specific protein factors.
Symbolic diagram of mRNA.
Symbolic diagram of an aminoacyl-tRNA.
Ribosomes of prokaryotes and eukaryotes.
Stages of Eukaryotic Translation
1. Activation of amino acids
2. Initiation
3. Elongation
4. Termination
Activation of Amino Acid
Activation of amino acid takes place in the cytosol.
In activation of amino acid each of the 20 amino
acids, is covalently attached to their respective tRNA,
at the expense of ATP, by aminoacyl tRNA synthases
(AAS) enzyme
Activation of amino acid.
(AAS: aminoacyl-tRNA synthetase)
Initiation
1. Ribosomal dissociation
2. Formation of 43S pre-initiation complex.
3. Formation of 48S initiation complex
4. Formation of 80S initiation complex.
Ribosomes have three sites:
1. Aminoacyl site (A site)
2. Peptidyl (P) site
3. Exit (E) site.
A and P sites bind to aminoacyl–tRNAs,
E site binds only to uncharged tRNAs
40S subunit of ribosome binds two initiation
factors elF3 and elF-1A and dissociates 80S into
two subunits 40S and 60S
Formation of 43S pre-initiation complex
Binding of GTP with initiation factors elF2.
GTP-elF2 binds to Met-tRNA Met
GTP-elF2-tRNA a ternary complex binds to 40S
ribosomal subunit to form 43S pre-initiation complex
43S pre-initiation complex is stabilized by association
with elF3 and elF-1A
In prokaryotes, the first amino acid methionine is modified
by formylation of its amino group to N-formyl-methionine
(f Met) and initiator tRNA is fmet-tRNAfMet.
Formation of 48S initiation complex
Binding of mRNA to the 43S pre-initiation complex
forms 48S initiation complex.
The association of mRNA with 43S initiation
complex requires cap binding protein (CBP) or elF4
and ATP
Formation of 80S initiation complex
Combination of the 48S initiation complex with 60S
ribosomal subunit forms 80S initiation complex.
Hydrolysis of the GTP bound to elF2, by elF5
Release of the initiation factors bound to the 48S
initiation complex.
At this stage, the Met-tRNAMet, is on the P site of the
ribosome so that its anticodon pairs with the initiating
AUG codon on the mRNA and is now ready for the
elongation process
Elongation
Elongation can be divided into following steps
1. Binding of aminoacyl-tRNA specified by the next codon
in the mRNA to the A site
2. Formation of peptide bond between amino acids
present in P and A site of ribosome
3. Translocation of the ribosome along the mRNA
4. Elimination of deacylated tRNA from the P and E sites.
Formation of peptide bond at site of the ribosome.
Termination
Elongation steps are repeated until one of the three
(UAA, UAG, UGA) termination or nonsense codon of
mRNA appears in A site.
Once the ribosome reaches a termination codon,
releasing factors are capable of recognizing the
termination signal present in the A site.
Prokaryotes have three release factors, RF-1, RF-2, and
RF-3 but eukaryotes have only single release factor, eRF.
Releasing factors, GTP, and peptidyl transferase
hydrolyzes the peptidyl tRNA bond
This hydrolysis releases the polypeptide and tRNA from
P site.
Upon hydrolysis and release of polypeptide the
mRNA is then released from the ribosome.
Ribosome then dissociates into 60S and 40S
subunits, which are then recycled.
(POST-TRANSLATIONAL MODIFICATION)
Types of post-trans- lational modifications are
1 Amino Terminal and Carboxyl Terminal
Modifications
2 Loss of Signal Sequence
3 Attachment of Carbohydrate Side Chains
4. Modification of Individual Amino Acids
Glycosylation
Phosphorylation
Carboxylation
Hydroxylation
Methylation
Addition of prosthetic group
5. Proteolytic Processing
6. Formation of Disulfide Cross Links
Inhibitors Of Protein Synthesis
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