T.Y.B.
Sc CBCS Module 7 (2024-25) As per KBC NMU, Jalgaon
Translation
Introduction
• Translation takes place on ribosomes.
• Ribosomes can be thought of as moving protein-synthesizing machines.
• A ribosome attaches near the 51 end of an mRNA strand and moves
toward the 31 end, translating the codons as it goes.
• Synthesis begins at the amino end of the protein, and the protein is
elongated by the addition of new amino acids to the carboxyl end.
Steps of Translation
Translation can be conveniently divided into four stages:
1. the binding of amino acids to the tRNAs
2. initiation, in which the components necessary for translation are
assembled at the ribosome
3. elongation, in which amino acids are joined, one at a time, to the growing
polypeptide chain and
4. termination, in which protein synthesis halts at the termination codon and
the translation components are released from the ribosome.
The Binding of Amino Acids to Transfer RNAs
• The first stage of translation is the binding of tRNA molecules to their
appropriate amino acids.
• When linked to its amino acid, a tRNA delivers that amino acid to the
ribosome, where the tRNA’s anticodon pairs with a codon on mRNA.
• This process enables the amino acids to be joined in the order specified
by the mRNA.
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• A cell typically possesses 30 to 50 different tRNAs, and, collectively,
these tRNAs are attached to the 20 different amino acids.
• Each tRNA is specific for a particular kind of amino acid.
• All tRNAs have the sequence CCA at the 3 end, and the carboxyl group
(COO) of the amino acid is attached to the 2- or 3- hydroxyl group of the
adenine nucleotide at the end of the tRNA.
• The key to specificity between an amino acid and its tRNA is a set of
enzymes called aminoacyl-tRNA synthetases.
• A cell has 20 different aminoacyl-tRNA synthetases, one for each of the
20 amino acids.
• Each synthetase recognizes a particular amino acid, as well as all the
tRNAs that accept that amino acid.
• Recognition of the appropriate amino acid by a synthetase is based on the
different sizes, charges, and R groups of the amino acids.
• The attachment of a tRNA to its appropriate amino acid (termed tRNA
charging) requires energy, which is supplied by adenosine triphosphate
(ATP)
amino acid+tRNA+ATP → aminoacyl-tRNA+AMP+PPi
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Initiation of Translation
The second stage in the process of protein synthesis is initiation. During
initiation, all the components necessary for protein synthesis assemble:
(1) mRNA; (2) the small and large subunits of the ribosome; (3) a set of three
proteins called initiation factors; (4) initiator tRNA with N-formylmethionine
attached (fMet-tRNAfMet); and (5) guanosine triphosphate (GTP).
Initiation comprises three major steps.
First, mRNA binds to the small subunit of the ribosome.
Second, initiator tRNA binds to the mRNA through base pairing between
the codon and anticodon.
Third, the large ribosome joins the initiation complex.
An mRNA molecule can bind to the small ribosome subunit only when the
subunits are separate.
Initiation factor 3 (IF-3) binds to the small subunit of the ribosome
preventing the large subunit from binding during initiation. The sequence
covered by the ribosome is from 30 to 40 nucleotides long and includes
the AUG initiation codon.
Within the ribosome-binding site is the Shine-Dalgarno consensus
sequence which is complementary to a sequence of nucleotides at the 3 1
end of 16S rRNA.
This allows the small subunit of the ribosome to attach to the mRNA and
positions the ribosome directly over the initiation codon
The initiator fMet-tRNAfMet attaches to the initiation codon. This step
requires initiation factor 2 (IF-2), which forms a complex with GTP. A
third factor, initiation factor 1 (IF-1), enhances the dissociation of the
large and small ribosomal subunits.
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At this point, the initiation complex consists of (1) The small subunit of
the ribosome; (2) the mRNA; (3) the initiator tRNA with its amino acid
(fMet-tRNAfMet); (4) one molecule of GTP; and (5) IF-3, IF-2, and IF-1.
These components are collectively known as the 30S initiation complex.
In the final step of initiation, IF-3 dissociates from the small subunit,
allowing the large subunit of the ribosome to join the initiation complex.
The molecule of GTP (provided by IF-2) is hydrolyzed to guanosine
diphosphate (GDP), and IF-1 and IF-2 depart. When the large subunit has
joined the initiation complex, it is called the 70S initiation complex.
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Elongation of Translation
The next stage in protein synthesis is elongation, in which amino acids
are joined to create a polypeptide chain.
Elongation requires (1) the 70S complex (2) tRNAs charged with their
amino acids; (3) several elongation factors (EF-Ts, EF-Tu, and EF-G);
and (4) GTP.
A ribosome has three sites that can be occupied by tRNAs; the
aminoacyl, or A, site, the peptidyl, or P, site, and the exit, or E, site The
initiator tRNA immediately occupies the P site (the only site to which the
fMet-tRNAfMet is capable of binding), but all other tRNAs first enter the
A site. After initiation, the ribosome is attached to the mRNA, and fMet-
tRNAfMet is positioned over the AUG start codon in the P site; the
adjacent A site is unoccupied
Elongation occurs in three steps.
The first step is the delivery of a charged tRNA (tRNA with amino acid)
to the A site.
This requires the presence of elongation factor Tu (EF-Tu), elongation
factor Ts (EF-Ts), and GTP. EF-Tu first joins with GTP and then binds
to a charged tRNA to form a three-part complex.
This three-part complex enters the A site of the ribosome, where the
anticodon on the tRNA pairs with the codon on the mRNA.
After the charged tRNA is in the A site, GTP is cleaved to GDP, and the
EF-Tu–GDP complex is released. Factor EF-Ts regenerates EF-Ts–GDP
to EF-Tu–GTP.
• The second step of elongation is the creation of a peptide bond between
the amino acids that are attached to tRNAs in the P and A sites.
• The formation of this peptide bond releases the amino acid in the P site
from its tRNA.
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• The activity responsible for peptide bond formation in the ribosome is
referred to as peptidyl transferase. This catalytic activity is a property of
the rRNA in the large subunit of the ribosome.
• This rRNA acts as a ribozyme
• The third step in elongation is translocation, the movement of the
ribosome down the mRNA in the 51 →31 direction.
• This step positions the ribosome over the next codon & requires
elongation factor G (EF-G) and GTP.
• The tRNAs in the P and A site are still attached to the mRNA through
codon– anticodon pairing. So they do not move with the ribosome as it
translocates.
• The ribosome shifts so that the tRNA that previously occupied the P site
now occupies the E site, from which it moves into the cytoplasm to be
recharged with another amino acid.
• Translocation also causes the tRNA that occupied the A site to be in the P
site, leaving the A site open.
• Thus, the progress of each tRNA through the ribosome during elongation
can be summarized as follows:
• cytoplasm → A site → P site → E site → : cytoplasm. The initiator tRNA
is an exception: it attaches directly to the P site and never occupies the A
site.
• After translocation, the A site of the ribosome is empty and ready to
receive the tRNA specified by the next codon.
• The elongation cycle repeats itself: a charged tRNA and its amino acid
occupy the A site, a peptide bond is formed between the amino acids in
the A and P sites, and the ribosome translocates to the next codon.
• Throughout the cycle, the polypeptide chain remains attached to the
tRNA in the P site.
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Termination of Translation
• Protein synthesis terminates when the ribosome translocates to a
termination codon.
• Because there are no tRNAs with anticodons complementary to the
termination codons, no tRNA enters the A site of the ribosome when a
termination codon is encountered.
• Instead, proteins called release factors bind to the ribosome.
• E. coli has three release factors—RF1, RF2, and RF3.
• Release factor 1 recognizes the termination codons UAA and UAG, and
RF2 recognizes UGA and UAA. Release factor 3 forms a complex with
GTP and binds to the ribosome.
• The release factors then promote the cleavage of the tRNA in the P site
from the polypeptide chain; in the process, the GTP that is complexed to
RF3 is hydrolyzed to GDP
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