Translation-An Overview: Professor (DR.) Namrata Chhabra Biochemistry For Medics - Lecture Notes WWW - Namrata.co

Translation- An overview
By- Professor (Dr.) Namrata Chhabra
Biochemistry For Medics- Lecture Notes

www.namrata.co
Biochemistry For Medics
07/12/15
Translation-Introduction
The pathway of protein synthesis is
called Translation because the

language of the nucleotide sequence
on the mRNA is translated into the
language of the amino acid sequence.
The m RNA is translated from its
5end to its 3end , producing a
protein synthesized from its amino
terminal end to its carboxyl terminal
end.
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Components required for

Translation Amino acids
Transfer RNA
Messenger RNA
Aminoacyl t RNA synthetase
Functionally competent ribosomes
Protein factors
ATP and GTP as a source of energy
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Steps of Protein
Synthesis
The process of protein synthesis is
divided into 3 stages- Initiation
- Elongation
- Termination
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Initiation
Initiation of the protein synthesis involves
the assembly of the components of the

translation system before the peptide
bond formation occurs. These components
include Two ribosomal subunits
m RNA, Aminoacyl tRNA specified by the
codons in the message ,GTP and initiation
factors that facilitate the assembly of this
initiation complex.
In prokaryotes three initiation factors are
known (IF-1,IF-2 and IF-3) while in
eukaryotes there are at least 9, designated
as e IF, to indicate the eukaryotic origin.
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Ribosomes
Ribosomes are large complexes of protein and
r RNA. They consist of two subunits- one large
and one small whose relative sizes are

generally given in terms of their
sedimentation coefficients or S (Svedberg)
values.
The S values are determined by the shape as
well as by the molecular mass, their
numerical values are strictly not additive.
The prokaryotic 50S and 30S ribosomal
subunits together form a ribosome with an S
value of 70.
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Structure of Ribosome
A and P sites on
Ribosome
The ribosome has two binding sites for t RNA
molecules, each of which extends over both
subunits. Together they cover the neighboring
codons.During translation ,
The A site binds an incoming Aminoacyl t RNA as
directed by the Codon currently occupying the site.
This Codon specifies the next amino acid to be
added to the growing peptide chain.
The P site codon is occupied by the peptidyl-t RNA.
This
t RNA carries the chain of amino acids that has already
been synthesized.
An E site is also there that is occupied by the empty
t RNA that is about to exit the ribosome
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Transfer RNA
At least one specific type of t RNA is
required per amino acid.
Two sites are important, one Amino acid
attachment site the other is Anticodon
site.
Because of their ability to carry a
specific amino acid and to recognize
the codons for that amino acid , t RNA
are called the adapter molecules.
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10
Structure of Transfer
RNA
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InitiationThe small ribosomal subunit binds to Initiation Factor 3

(IF3).
The small subunit/IF3 complex binds to the

mRNA. Specifically, it binds to the
sequence AGGAGG, known as the
Shine-Delgarno sequence, which is found in
all prokaryotic mRNAs.
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13
Meanwhile, the fmet tRNA binds to

Initiation Factor 2 (IF2), which promotes
binding of the tRNA to the start codon.
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The small subunit/IF3 complex scans along the mRNA until

it encounters the start codon. The tRNA/IF2 complex also
binds to the start codon. This complex of the small
ribosomal subunit, IF3, initiator tRNA, and IF2 is
called the initiation complex.
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At this point, the large ribosomal subunit joins

in. A molecule of GTP is hydrolyzed, and the
initiation factors are released. The ribosomal
complex is now ready for protein synthesis.
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When the ribosome is assembled, two tRNA binding sites

are created; these are designated 'P' and 'A' (P stands
for peptidyl, A stands for amino acyl). The initiator
tRNA is in the P site, and the A site will be filled by
the tRNA with the anticodon that is complementary to
the codon next to the start. (In this case, it is the
tRNA that binds proline.)
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When the second tRNA base pairs with the

appropriate codon in the mRNA, an enzyme called
peptidyl transferase catalyzes the formation of a
peptide bond between the two amino acids present
(while breaking the bond between fmet and its
tRNA).This activity is intrinsic to the 23S r RNA
found in the large subunit. Since the r RNA
catalyzes this process , it is referred to as the
Ribozyme
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ElongationAt this point, the whole ribosome shifts over one

codon. This shift requires several elongation factors
(not shown) and energy from the hydrolysis of GTP. The
result of the shift is that the uncharged tRNA that was
in the P site is ejected, and the tRNA that was in the
A site is now in the P site. The A site is free to
accept the tRNA molecule with the appropriate anticodon
for the next codon in the mRNA.
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The next tRNA base pairs with the next codon, and
peptidyl transferase catalyzes the formation of a
peptide bond between the new amino acid and the growing
peptide chain.
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Elongation (Contd.)
Once again, the ribosome shifts over, so that the
uncharged tRNA is expelled, and the tRNA with the
peptide chain occupies the P site. (This is why this
site is called the 'peptidyl' site - after the shift,
it contains the tRNA with the growing peptide chain.
The other site will accept a tRNA with an amino acid,
hence the name 'aminoacyl' site) .
The process of shifting and peptide bond formation
continues over and over until a termination codon is
encountered. The elongation process is fairly rapid,
with prokaryotic ribosomes able to add 15 amino acids
to the growing polypeptide every second.
The process is also relatively error-free. Only one
mistake is made every 10,000 amino acids. For large
proteins of 1000 amino acids, that would mean one wrong
amino acid in every 10 polypeptides.
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Elongation (Contd.)
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Termination-
When a termination codon enters the A site,

translation halts. This is because there is no
tRNA with an anti codon that is complementary to
any of the stop codons.
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The release factor causes the translation complex

to fall apart, and cleaves the polypeptide from
the final tRNA. The polypeptide product is now
free to function in the cell. The mRNA molecule is
now available to be translated again.
Very often, more than one ribosome will translate
a single mRNA at the same time. One ribosome will
initiate translation, and after it moves down the
mRNA a bit, another ribosome will initiate, then
another, and so on.
The structure consisting of multiple ribosomes
translating a single mRNA molecule is called a
polysome. Eventually, the mRNA is degraded, and
translation of that particular message will cease.
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Termination
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Eukaryotic Translation
Eukaryotic translation is very similar overall to prokaryotic translation.
There are a few notable differences, These include the followings:
Eukaryotic mRNAs do not contain a Shine-Delgarno sequence.
Instead, ribosomal subunits recognize and bind to the 5' cap of
eukaryotic mRNAs. In other words, the 5' cap takes the place of the
Shine-Delgarno sequence.
Eukaryotes do not use formyl methionine as the first amino acid in
every polypeptide; ordinary methionine is used.
Eukaryotes do have a specific initiator tRNA, however.
Eukaryotic translation involves many more protein factors than
prokaryotic translation (For example, eukaryotic initiation involves at
least 10 factors, instead of the 3 in prokaryotes.)
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Inhibitors of protein
synthesis
The tetracyclines (tetracycline, doxycycline,
demeclocycline, minocycline, etc.) block

bacterial translation by binding reversibly to
the 30S subunit and distorting it in such a way
that the anticodons of the charged tRNAs
cannot align properly with the codons of the
mRNA.
Puromycin structurally binds to the amino acyl
t RNA and becomes incorporated into the
growing peptide chain thus causing inhibition of
the further elongation.
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Inhibitors of protein
synthesis
Choramphenicol inhibits prokaryotic
peptidyl Transferase
Clindamycin and Erythromycin bind
irreversibly to a site on the 50 s subunit
of the bacterial ribosome thus inhibit
translocation.
Diphtheria toxin inactivates the
eukaryotic elongation factors thus
prevent translocation.
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Post Translational
Modifications
The newly synthesized protein is modified to
become functionally active. The various post
translational modifications are as follows-Trimming
-Covalent Alterations
a)Phosphorylation
b) Glycosylation
c)Hydroxylation
d)Gamma carboxylation
e) Isoprenylation
-Protein degradation
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Post Translational
Modifications
Trimming removes excess amino acids.
Phosphorylation may activate or inactivate the
protein
Glycosylation targets a protein to become a part of
the plasma membrane , or lysosomes or be
secreted out of the cell
Hydroxylation such as seen in collagen is required
for acquiring the three dimensional structure and for
imparting strength
Defective proteins or destined for turn over are
marked for destruction by attachment of a
Ubiquitin protein. Proteins marked in this way are
degraded by a cellular component known as the
Proteasome.
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Translation: Summary of Key

Points
Translation is the synthesis of a polypeptide using the
information encoded in an mRNA molecule. The process

involves mRNA, tRNA, and ribosomes.
tRNA has a unique structure that exposes an anticodon,
which binds to codons in an mRNA, and an opposite end
that binds to a specific amino acid. Binding of an amino
acid to a tRNA is carried out by an enzyme called
aminoacyl tRNA synthetase in a process called charging.
Translation consists of three basic steps: initiation,
elongation, and termination. Initiation involves the
formation of the ribosome/mRNA/initiator tRNA complex.
Elongation is the actual synthesis of the polypeptide
chain, by formation of peptide bonds between amino
acids.
Termination dissociates the translation complex and
releases the finished polypeptide chain. Each of these
steps requires the activity of a specific set of protein
factors in addition to the ribosome, tRNA, and mRNA.
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Summary of Translation
07/12/15
32

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Translation- An overview

By- Professor (Dr.) Namrata Chhabra

Biochemistry For Medics- Lecture Notes

called Translation because the

Components required for

Biochemistry For Medics

Biochemistry For Medics

the assembly of the components of the

and one small whose relative sizes are

Biochemistry For Medics

Biochemistry For Medics

InitiationThe small ribosomal subunit binds to Initiation Factor 3

The small subunit/IF3 complex binds to the

Biochemistry For Medics

Meanwhile, the fmet tRNA binds to

Biochemistry For Medics

The small subunit/IF3 complex scans along the mRNA until

Biochemistry For Medics

At this point, the large ribosomal subunit joins

Biochemistry For Medics

When the ribosome is assembled, two tRNA binding sites

Biochemistry For Medics

When the second tRNA base pairs with the

Biochemistry For Medics

ElongationAt this point, the whole ribosome shifts over one

Biochemistry For Medics

Biochemistry For Medics

Biochemistry For Medics

When a termination codon enters the A site,

Biochemistry For Medics

The release factor causes the translation complex

Biochemistry For Medics

Biochemistry For Medics

Biochemistry For Medics

demeclocycline, minocycline, etc.) block

Translation: Summary of Key

information encoded in an mRNA molecule. The process

Biochemistry For Medics

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