Transcription Ribosomes are the structures where polypeptides (proteins) are

built. They are made up of protein and RNA (ribosomal RNA, or

Transcription is the first step of gene expression. During this
rRNA). Each ribosome has two subunits, a large one and a small one,
process, the DNA sequence of a gene is copied into RNA. Before
which come together around an mRNA. The ribosome provides a set
transcription can take place, the DNA double helix must unwind
of handy slots where tRNAs can find their matching codons on the
near the gene that is getting transcribed. The region of opened-up
mRNA template and deliver their amino acids. These slots are called
DNA is called a transcription bubble. Transcription uses one of the
the A, P, and E sites. Not only that, but the ribosome also acts as an
two exposed DNA strands as a template; this strand is called the
enzyme, catalyzing the chemical reaction that links amino acids
template strand. The RNA product is complementary to the
together to make a chain.
template strand and is almost identical to the other DNA strand,
called the non-template (or coding) strand. The site on the DNA Transcription in Prokaryotes
from which the first RNA nucleotide is transcribed is called the +1
site or the initiation site. Nucleotides that come before the initiation
site are given negative numbers and said to be upstream.
Nucleotides that come after the initiation site are marked with
positive numbers and said to be downstream.

Translation

Two types of molecules with key roles in translation are tRNAs and
ribosomes.

Transfer RNAs (tRNAs)

Transfer RNAs, or tRNAs, are molecular "bridges" that connect transcription is an enzymatic process. the mechanism of
mRNA codons to the amino acids they encode. One end of each transcription completes in three major steps
tRNA has a sequence of three nucleotides called an anticodon,
1. Initiation:
which can bind to specific mRNA codons. The other end of the tRNA
carries the amino acid specified by the codons.  closed complex formation
 Open complex formation
There are many different types of tRNAs. Each type reads one or a
 Tertiary complex formation
few codons and brings the right amino acid matching those codons.
2. Elongation
Ribosomes
3.Termination:
  Rho- dependent the time, or infrequently. Although promoters vary among
 Rho-independent prokaryotic genomes, a few elements are conserved. At the -10 and
-35 regions upstream of the initiation site, there are two promoter
1. Initiation
consensus sequences, or regions that are similar across all
The transcription is initiated by RNA polymerase holoenzyme from a promoters and across various bacterial species. Binding of RNA
specific point called promotor sequence. The core enzyme bind to polymerase holoenzyme to the promotor sequence form closed
specific sequence on template DNA strand called promotor. The complex
binding of core polymerase to promotor is facilitates and specified
The -10 consensus sequence, called the -10 region, is TATAAT. The
by sigma (σ) factor.
-35 sequence, TTGACA, is recognized and bound by σ. Once this
interaction is made, the subunits of the core enzyme bind to the
site. The A–T-rich -10 region facilitates unwinding of the DNA
template, and several phosphodiester bonds are made. The
transcription initiation phase ends with the production of abortive
transcripts, which are polymers of approximately 10 nucleotides
that are made and released. So that open complex is formed. This
changing from closed complex to open complex is called
isomerization.

These subunits assemble every time a gene is transcribed, and they RNA polymerase starts synthesizing nucleotide. It doesnot require
disassemble once transcription is complete. Each subunit has a the help of primase. If the enzyme synthesize short RNA molecules
unique role; the two α-subunits are necessary to assemble the of less than 10 bp, it does not further elongates which is called
polymerase on the DNA; the β-subunit binds to the ribonucleoside abortive initiation. When the RNA polymerase manage to synthesize
triphosphate that will become part of the nascent mRNA molecule; RNA more than 10 bp long, it eject the σ 3.2 region and RNA further
and the β‘ subunit binds the DNA template strand. The fifth subunit, elongates and exit from RNA exit channel. This is the formation of
σ, is involved only in transcription initiation. Without σ, the core tertiary complex.
enzyme would transcribe from random sites

A promoter is a DNA sequence onto which the transcription

machinery binds and initiates transcription. In most cases,
promoters exist upstream of the genes they regulate. The specific
sequence of a promoter is very important because it determines
whether the corresponding gene is transcribed all the time, some of
 The synthesized RNA exit from RNA exit channel. The synthesized
RNA is proof reads by Hydrolytic editing. For this the polymerase
back track by one or more nucleotide and cleave the RNA removing
the error and synthesize the correct one. The Gre factor enhance
this proof reading process. Pyrophospholytic editing another
mechanism of removing altered nucleotide.

Proof reading

Pyrophosphorolytic editing：The enzyme uses its active site ,in a

simple back-reaction, to catalyze the removal of an incorrectly
inserted ribonucleotide, by reincorporation of PPi. When removing
bases, the enzyme spends longer hovering over mismatches than
matches, so removes the former more frequently.
2. Elongation Hydrolytic editing: The polymerase backtracks by one or more
nucleotides and cleaves the RNA product, removing the error-
The transcription elongation phase begins with the release of the
containing sequence. The Gre factors enhances the hydrolytic
σsubunit from the polymerase. The dissociation of σ allows the core
editing function,serves as elongation stimulating factors as well.
RNA polymerase enzyme to proceed along the DNA template,
synthesizing mRNA in the 5’ to 3’ direction at a rate of
approximately 40 nucleotides per second. As elongation proceeds, 3. Termination
the DNA is continuously unwound ahead of the core enzyme and
rewound behind it. Since the base pairing between DNA and RNA is Rho independent:
not stable enough to maintain the stability of the mRNA synthesis In this mechanism, transcription is terminated due to specific
components, RNA polymerase acts as a stable linker between the sequence in terminator DNA.The terminator DNA contains invert
DNA template and the nascent RNA strands to ensure that repeat which cause complimentary pairing as transcript RNA form
elongation is not interrupted prematurely. hair pin structure.This invert repeat is followed by larger number of
Free NTPs are added sequentially to the 3’-OH of the nascent RNA TTTTTTTT(~8 bp) on template DNA. The uracil appear in RNA. The
strand. load of hair pin structure is not tolerated by A=U base pair so the
RNA get separated from transcription bubble.
(NMP)n + NTP (NMP)n+1 + PP
 Steps in translation:

Activation of aminoacids:

The activation of amino acids take place in cytosol. The activation of

amino acids is catalyzed by their aminoacyl tRNA synthetases.

All the 20 amino acids are activated and bound to 3’ end of their
specific tRNA in the presence of ATP and Mg++.

The N-formylated methionine is chain initiating amino acid in

bacteria whereas methionine is chain initiating amino acid in
eukaryotes.

Methionine is activated by methionyl-tRNA synthetase. For N-

formylmethionine two types of tRNA are used ie. tRNAmet and
tRNAfmet.
Rho depended:
Similarly, all 20 aminoacids are activated (amino acyl-AMP enzyme
In this mechanism, transcription is terminated by rho (ρ) protein.It is complex) and then bound to their specific tRNA forming Aminoacyl
ring shaped single strand binding ATpase protein. The rho protein tRNA.
bind the single stranded RNA as it exit from polymerase enzyme
complex and hydrolyse the RNA from enzyme complex. The rho
protein does not bind to those RNA whose protein is being
translated. Rather it bind to RNA after translation. In bacteria
transcription and translation occur simultaneously so the rho
protein bind the RNA after translation has completed but
transcription is still ON.

Initiation:
Translation in prokaryotes
In the first step, initiation factor-3 (IF-3) binds to 30S ribosomal unit.
It is the process of synthesis of protein by encoding information on
mRNA. Protein synthesis requires mRNA, tRNA, aminoacids,
ribosome and enzyme aminoacyl tRNA synthase
Then mRNA binds to 30S ribosomal subunit in such a way that AUG iii. Ribosome translocation:
codon lie on the peptidyl (P) site and the second codon lies on
After peptide bond formation ribosome moves one codon ahead
aminoacyl (A) site.
along 5’-3’ direction on mRNA, so that dipeptide-tRNA appear on P-
The tRNA carrying formylated methionine ie. FMet–tRNA FMet is site and next codon appear on A-site.
palced at P-site. This specificity is induced by IF-2 with utilization of
The uncharged tRNA exit from ribosome and enter to cytosol.
GTP. The IF-1 prevent binding of FMet–tRNAF Met is in A-site.
The ribosomal translocation requires EF-G-GTP (translocase
Shinedalgrno sequence in the mRNA guide correct positioning of
enzyme) which change the 3D structure of ribosome and catalyze
AUG codon at P-site of 30S ribosome.
5’-3’ movement.
After binding of FMet–tRNAFMet on P-site, IF-3, IF-2 and IF-1 are
The codon on A-site is now recognized by other aminoacyl-tRNA as
released so that 50S ribosomal unit bind with 30S forming 70S
in previous.
sibosome. The exit site is located in 50S.
The dipeptide on P-site is transferred to A-site forming tripeptide.
3. Elongation:
This process continues giving long polypeptide chain of aminoacids.
i. Binding of AA-tRNA at A-site:
4. Termination:
The 2nd tRNA carrying next aminoacid comes into A-site and
recognizes the codon on mRNA. This binding is facilitated by EF-TU The peptide bond formation and elongation of polypeptide
and utilizes GTP. continues until stop codon appear on A-site.
After binding, GTP is hydrolysed and EF-TU-GDP is releasd If stop codon appear on A-site it is not recognized by t-RNA carrying
amino acids because stop codon donot have anticodon on mRNA.
EF=TU-GDP then and enter into EF-TS cycle.
The stop codon are recognized by next protein called release factor
ii. Peptide bond formation:
(Rf-1, RF-2 and RF-3) which hydrolyses and cause release of all
The aminoacid present in t-RNA of P-site ie Fmet is transferred to t- component ie 30s, 50S, mRNA and polypeptide separates.
RNA of A-site forming peptide bond. This reaction is catalyzed by
RF-1 recognisaes UAA and UAg while RF-2 recognises UAA and UGA
peptidyltransferase.
while RF-3 dissociate 30S and 50S subunits.
Now, the t-RNA at P-site become uncharged
In case of eukaryotes only one release actor eRF causes dissociation.
Post translation modification:

The newly formed polypeptide may not be biologically functional so

it undergoes several folding and processing known as post
translation modification.

1. Amino terminal and carboxyl terminal modification: The N-

formylmethionine in case of bacteria is removed from polypeptide
chain and some carboxyl terminal are also removed by enzymatic
action to make functional protein. In case of eukaryotic protein,
amino terminal is N- acetylated.

2. Loss of signal sequences: In some protein the amino terminal end

is cleaved by specific peptidase so that protein loss its signaling
property.

3. Modification of individual aminoacids: The aminoacids may be

phosphorylated, acetylated for modification

4. Attachment of carbohydrate side chain: Carbohydrate side chain

is added to make protein functional. Eg, glycoprotein. Lipoprotein

5. Addition of isoprenyl group: In some protein, isoprenyl group is

added so to make protein active

Various protein factors involved in protein synthesis

Transcription in eukaryotes several types of regulatory RNAs, including microRNAs (miRNAs)
and long-coding RNAs (lncRNAs).
Eukaryotic transcription is carried out in the nucleus of the cell by
one of three RNA polymerases, depending on the RNA being RNA polymerase III is also located in the nucleus. This polymerase
transcribed, and proceeds in three sequential stages: transcribes a variety of structural RNAs that includes the 5S pre-
rRNA, transfer pre-RNAs (pre-tRNAs), and small nuclear pre-RNAs.
1. Initiation
The tRNAs have a critical role in translation: they serve as the
2. Elongation
adaptor molecules between the mRNA template and the growing
3. Termination.
polypeptide chain. Small nuclear RNAs have a variety of functions,
The Three Eukaryotic RNA Polymerases (RNAPs) including “splicing” pre-mRNAs and regulating transcription factors.
Not all miRNAs are transcribed by RNA Polymerase II, RNA
The features of eukaryotic mRNA synthesis are markedly more Polymerase III transcribes some of them.
complex those of prokaryotes. Instead of a single polymerase
comprising five subunits, the eukaryotes have three polymerases Initiation of Transcription in Eukaryotes
that are each made up of 10 subunits or more. Each eukaryotic
Unlike the prokaryotic RNA polymerase that can bind to a DNA
polymerase also requires a distinct set of transcription factors to
template on its own, eukaryotes require several other proteins,
bring it to the DNA template.
called transcription factors, to first bind to the promoter region and
RNA polymerase I is located in the nucleolus, a specialized nuclear then help recruit the appropriate polymerase. The completed
substructure in which ribosomal RNA (rRNA) is transcribed, assembly of transcription factors and RNA polymerase bind to the
processed, and assembled into ribosomes. The rRNA molecules are promoter, forming a transcription pre-initiation complex (PIC).
considered structural RNAs because they have a cellular role but are
The most-extensively studied core promoter element in eukaryotes
not translated into protein. The rRNAs are components of the
is a short DNA sequence known as a TATA box, found 25-30 base
ribosome and are essential to the process of translation. RNA
pairs upstream from the start site of transcription. Only about 10-
polymerase I synthesizes all of the rRNAs except for the 5S rRNA
15% of mammalian genes contain TATA boxes, while the rest
molecule.
contain other core promoter elements, but the mechanisms by
RNA polymerase II is located in the nucleus and synthesizes all which transcription is initiated at promoters with TATA boxes is well
protein-coding nuclear pre-mRNAs. Eukaryotic pre-mRNAs undergo characterized.
extensive processing after transcription, but before translation. RNA
The TATA box, as a core promoter element, is the binding site for a
polymerase II is responsible for transcribing the overwhelming
transcription factor known as TATA-binding protein (TBP), which is
majority of eukaryotic genes, including all of the protein-encoding
itself a subunit of another transcription factor: Transcription Factor
genes which ultimately are translated into proteins and genes for
II D (TFIID). After TFIID binds to the TATA box via the TBP, five more
transcription factors and RNA polymerase combine around the Eukaryotic pre-mRNA processing
TATA box in a series of stages to form a pre-initiation complex. One
 When an RNA transcript is first made in a eukaryotic cell, it
transcription factor, Transcription Factor II H (TFIIH), is involved in
is considered a pre-mRNA and must be processed into a
separating opposing strands of double-stranded DNA to provide the
messenger RNA (mRNA).
RNA Polymerase access to a single-stranded DNA template.
 A 5' cap is added to the beginning of the RNA transcript, and
However, only a low, or basal, rate of transcription is driven by the
a 3' poly-A tail is added to the end.
pre-initiation complex alone. Other proteins known as activators
and repressors, along with any associated coactivators or  In splicing, some sections of the RNA transcript (introns) are
corepressors, are responsible for modulating transcription rate. removed, and the remaining sections (exons) are stuck back
Activator proteins increase the transcription rate, and repressor together.
proteins decrease the transcription rate.  Some genes can be alternatively spliced, leading to the
production of different mature mRNA molecules from the
same initial transcript.

In bacteria, RNA transcripts are ready to act as messenger RNAs and

get translated into proteins right away. In eukaryotes, things are a
little more complex. The molecule that's directly made by
transcription is called a pre-mRNA, reflecting that it needs to go
through a few more steps to become an actual messenger RNA
(mRNA). These are:

 Addition of a 5' cap to the beginning of the RNA

 Addition of a poly-A tail (tail of A nucleotides) to the end of
the RNA
 Chopping out of introns, or "junk" sequences, and pasting
together of the remaining, good sequences (exons)

5' cap and poly-A tail

Both ends of a pre-mRNA are modified by the addition of chemical

groups. The group at the beginning (5' end) is called a cap, while the
group at the end (3' end) is called a tail. Both the cap and the tail
protect the transcript and help it get exported from the nucleus and
translated on the ribosomes (protein-making "machines") found in Splicing needs to precise and consistent. This careful cutting and
the cytosol pasting is performed by the spliceosome, an enzyme complex made
of protein and small RNAs. Most introns contain marker sequences
The 5’ cap is added to the first nucleotide in the transcript during
at both of their ends, which are recognized by the small RNAs and
transcription. The cap is a modified guanine (G) nucleotide, and it
direct the spliceosome to remove the intron. Once the intron has
protects the transcript from being broken down. It also helps the
been cut out, the spliceosome will "glue" (ligate) the flanking exons
ribosome attach to the mRNA and start reading it to make a protein.
together.
When a sequence called a polyadenylation signal shows up in an
Elongation in eukaryotes
RNA molecule during transcription, an enzyme chops the RNA in
two at that site. Another enzyme adds about 100-200 adenine (A) RNA Polymerase II is a complex of 12 protein subunits. Specific
nucleotides to the cut end, forming a poly-A tail. The tail makes the subunits within the protein allow RNA Polymerase II to act as its
transcript more stable and helps it get exported from the nucleus to own helicase, sliding clamp, single-stranded DNA binding protein, as
the cytosol. well as carry out other functions. Consequently, RNA Polymerase II
does not need as many accessory proteins to catalyze the synthesis
RNA splicing
of new RNA strands during transcription elongation as DNA
The third big RNA processing event that happens is RNA splicing. In Polymerase does to catalyze the synthesis of new DNA strands
RNA splicing, specific parts of the pre-mRNA, called introns are during replication elongation.
recognized and removed by a protein-and-RNA complex called the
However, RNA Polymerase II does need a large collection of
spliceosome. Introns can be viewed as "junk" sequences that must
accessory proteins to initiate transcription at gene promoters, but
be cut out so the "good parts version" of the RNA molecule can be
once the double-stranded DNA in the transcription start region has
assembled.
been unwound, the RNA Polymerase II has been positioned at the
What are the "good parts"? The pieces of the RNA that are not +1 initiation nucleotide, and has started catalyzing new RNA strand
chopped out are called exons. The exons are pasted together by the synthesis, RNA Polymerase II clears or “escapes” the promoter
spliceosome to make the final, mature mRNA that is shipped out of region and leaves most of the transcription initiation proteins
the nucleus. behind.

A key point here is that it's only the exons of a gene that encode a All RNA Polymerases travel along the template DNA strand in the 3’
protein. Not only do the introns not carry information to build a to 5’ direction and catalyze the synthesis of new RNA strands in the
protein, they actually have to be removed in order for the mRNA to 5’ to 3’ direction, adding new nucleotides to the 3’ end of the
encode a protein with the right sequence. If the spliceosome fails to growing RNA strand.
remove an intron, an mRNA with extra "junk" in it will be made, and
a wrong protein will get produced during translation.
RNA Polymerases unwind the double stranded DNA ahead of them The protein-encoding, structural RNA, and regulatory RNA genes
and allow the unwound DNA behind them to rewind. As a result, transcribed by RNA Polymerse II lack any specific signals or
RNA strand synthesis occurs in a transcription bubble of about 25 sequences that direct RNA Polymerase II to terminate at specific
unwound DNA basebairs. Only about 8 nucleotides of newly- locations. RNA Polymerase II can continue to transcribe RNA
synthesized RNA remain basepaired to the template DNA. The rest anywhere from a few bp to thousands of bp past the actual end of
of the RNA molecules falls off the template to allow the DNA behind the gene. However, the transcript is cleaved at an internal site
it to rewind. before RNA Polymerase II finishes transcribing. This releases the
upstream portion of the transcript, which will serve as the initial
RNA prior to further processing (the pre-mRNA in the case of
RNA Polymerases use the DNA strand below them as a template to protein-encoding genes.) This cleavage site is considered the “end”
direct which nucleotide to add to the 3’ end of the growing RNA of the gene. The remainder of the transcript is digested by a 5′-
strand at each point in the sequence. The RNA Polymerase travels exonuclease (called Xrn2 in humans) while it is still being
along the template DNA one nucleotide at at time. Whichever RNA transcribed by the RNA Polymerase II. When the 5′-exonulease
nucleotide is capable of basepairing to the template nucleotide “catches up” to RNA Polymerase II by digesting away all the
below the RNA Polymerase is the next nucleotide to be added. Once overhanging RNA, it helps disengage the polymerase from its DNA
the addition of a new nucleotide to the 3′ end of the growing strand template strand, finally terminating that round of transcription.
has been catalyzed, the RNA Polymerase moves to the next DNA
In the case of protein-encoding genes, the cleavage site which
nucleotide on the template below it. This process continues until
determines the “end” of the emerging pre-mRNA occurs between
transcription termination occurs.
an upstream AAUAAA sequence and a downstream GU-rich
Termination sequence separated by about 40-60 nucleotides in the emerging
RNA. Once both of these sequences have been transcribed, a
The termination of transcription is different for the three different protein called CPSF in humans binds the AAUAAA sequence and a
eukaryotic RNA polymerases. protein called CstF in humans binds the GU-rich sequence. These
The ribosomal rRNA genes transcribed by RNA Polymerase I contain two proteins form the base of a complicated protein complex that
a specific sequence of basepairs (11 bp long in humans; 18 bp in forms in this region before CPSF cleaves the nascent pre-mRNA at a
mice) that is recognized by a termination protein called TTF-1 site 10-30 nucleotides downstream from the AAUAAA site. The
(Transcription Termination Factor for RNA Polymerase I.) This Poly(A) Polymerase enzyme which catalyzes the addition of a 3′
protein binds the DNA at its recognition sequence and blocks poly-A tail on the pre-mRNA is part of the complex that forms with
further transcription, causing the RNA Polymerase I to disengage CPSF and CstF.
from the template DNA strand and to release its newly-synthesized Translation in eukaryotes
RNA.
1. Site:

Translation occurs in the cytoplasm where the ribosomes are

located. Ribosomes are made of a small and large subunit which
surrounds the mRNA. In eukaryotic translation 80S ribosomes with
40S and 60S subunits are used. The mRNA is synthesized from DNA
only. In eukaryotes, there is single initiation and termination site.

2. Template:

This uses an mRNA sequence as a template to guide the synthesis of

a chain of amino acids that form a protein. Many types of
transcribed RNA, such as transfer RNA, ribosomal RNA, and small
nuclear RNA are not necessarily translated into an amino acid
sequence.

4. Factors Involved:

In eukaryotes, several factors are used in chain initiation such as (initialtion factor)
eIF2, eIF3, eIF4A, eIF4E, eIF4F and elF 4G. Two factors [EF-1 and EF-
2] are used in chain elongation. There is a single release factor RF
for recognition of three termination codons [UAA, UAG and UGA]. Initiation of Translation

5. Enzymes Involved: The initiation of translation in eukaryotes is complex, involving at

least ten eukaryotic initiation factors (eIFs). Some of the eIFs
In eukaryotes, two types of enzymes are used in translation. contain multiple (3-8) subunits. The process of translation initiation
Aminoacyl tRNA synthetase (an enzyme) catalyzes the bonding can be divided into four steps.
between specific tRNAs and the amino acids. The enzyme peptidyl
transferase connect A site and P site by forming a peptide bond [the Ribosomal dissociation.
nitrogen carbon bond] during elongation phase. (i) Formation of 43S preinitiation complex.
(ii) Formation of 48S initiation complex.
(iii) Formation of 80S initiation complex.
(iv) Ribosomal dissociation

The 80S ribosome dissociates to form 40S and 60S subunits.

Two initiating factors namely eIF3 and eIF-1A bind to the newly The ribosomal initiation complex scans the mRNA for the
formed 40S subunit, and thereby block its reassociation with 60S identification of appropriate initiation codon. 5c-AUG is the
subunit. For this reason, some workers name eIF-3 as anti- initiation codon and its recognition is facilitated by a specific
association factor. sequence of nucleotides surrounding it.

Formation of 43S preinitiation complex This marker sequence for the identification of AUG is called as Kozak
consensus sequence.
A ternary complex containing met-tRNAi and eIF-2 bound to GTP
attaches to 40S ribosomal subunit to form 43S preinitiation In case of prokaryotes the recognition sequence of initiation codon
complex. is referred to as Shine- Dalgarno sequence.

The presence of eIF-3 and eIF-1A stabilizes this complex (Note : -Formation of 80S initiation complex
Met-tRNA is specifically involved in binding to the initiation condon
48S initiation complex binds to 60S ribosomal subunit to form 80S
AUGs; hence the superscripti is used in mettRNAi).
initiation complex.
Formation of 48S initiation complex
The binding involves the hydrolysis of GTP (bound to eIF-2). This
The binding of mRNA to 43S preinitiation complex results in the step is facilitated by the involvement of eIF-5.
formation of 48S initiation complex through the intermediate 43S
As the 80S complex is formed, the initiation factors bound to 48S
initiation complex. This, however, involves certain interactions
initiation complex are released and recycled.
between some of the eIFs and activation of mRNA.
The activation of eIF-2 requires eIF-2B (also called as guanine
eIF-4F complex is formed by the association of eIF-4G, eIF-4A with
nucleotide exchange factor) and GTP.
eIF-4E.
The activated eIF-2 (i.e. bound to GTP) requires eIF2C to form the
The so formed eIF-4F (referred to as cap binding protein) binds to
ternary complex.
the cap of mRNA.
Regulation of initiation
Then elF-4A and elF-4B bind to mRNA and reduce its complex
structure. The eIF-4F, a complex formed by the assembly of three initiation
factors controls initiation, and thus the translation process.
This mRNA is then transferred to 43S complex.
eIF4E, a component of eIF-4F is primarily responsible for the
For the appropriate association of 43S preinitiation complex with
recognition of mRNA cap. And this step is the rate-limiting in
mRNA, energy has to be supplied by ATP.
translation.
Recognition of initiation codon:
eIF-2 which is involved in the formation of 43S preinitiation complex As the amino acid in the aminoacyl-tRNA is already activated, no
also controls protein biosynthesis to some extent. additional energy is required for peptide bond formation. The net
result of peptide bond formation is the attachment of the growing
Elongation of Translation
peptide chain to the tRNA in the A-site.
Ribosomes elongate the polypeptide chain by a sequential addition
Translocation
of amino acids. The amino acid sequence is determined by the order
of the codons in the specific mRNA. Elongation, a cyclic process As the peptide bond formation occurs, the ribosome moves to the
involving certain elongation factors (EFs), may be divided into three next codon of the mRNA (towards 3c-end). This process called
steps. translocation, basically involves the movement of growing peptide
chain from A-site to P-site.
(i) Binding of aminoacyl t-RNA to A-site.
(ii) Peptide bond formation. Translocation requires EF-2 and GTP.
(iii) Translocation.
GTP gets hydrolysed and supplies energy to move mRNA. EF-2 and
Binding of aminoacyl—tRNA to A-site GTP complex recycles for translocation.

The 80S initiation complex contains met-tRNAi in the P-site, and the In recent years, another site namely exit site (E-site) has been
A-site is free. identified in eukaryotes. The deacylated tRNA moves into the E-site,
from where it leaves the ribosome.
Another aminoacyl-tRNA is placed in the A-site. This requires
proper codon recognition on the mRNA and the involvement of In case of prokaryotes, the elongation factors are different, and they
elongation factor 1a (EF-Ia) and supply of energy by GTP. are EF-Tu, EF-Ts (in place of of EF-1a) and EF-G (instead of EF-2).

As the aminoacyl-tRNA is placed in the A-site, EF-1D and GDP are Incorporation of amino acids
recycled to bring another aminoacyl-tRNA.
It is estimated that about six amino acids per second are
Peptide bond formation incorporated during the course of elongation of translation in
eukaryotes. In case of prokaryotes, as many as 20 amino acids can
The enzyme peptidyltransferase catalyses the formation of peptide
be incorporated per second.Thus the process of
bond.
protein/polypeptide synthesis in translation occurs with great speed
The activity of this enzyme lies on 28S RNA of 60S ribosomal and accuracy.
subunit. It is therefore the rRNA (and not protein) referred to as
Termination of Translation
ribozyme that catalyses the peptide bond formation.
Termination is a simple process when compared to initiation and
elongation. After several cycles of elongation, incorporating amino
acids and the formation of the specific protein/ polypeptide
molecule, one of the stop or termination signals (UAA, UAG and
UCA) terminates the growing polypeptide.

The termination codons which act as stop signals do not have

specific tRNAs to bind. As theb termination codon occupies the
ribosomal A-site, the release factor namely eRF recognizes the stop
signal. eRF-GTP complex, in association with the enzyme
peptidyltransferase, cleaves the peptide bond between the
polypeptide and the tRNA occupying P-site. In this reaction, a water
molecule, instead of an amino acid is added. This hydrolysis releases
the protein and tRNA from the P-site. The 80S ribosome dissociates
to form 40S and 60S subunits which are recycled. The mRNA is also
released.

Eukaryotic initiation factor

Elongation factor for Prokaryote/Eukaryotic Translation  Removing the methionine- AUG code from the mature
chain.
 Adding a functional group to the amino acid or polypeptide
chain through phosphorylation, glycosylation, ubiquitination
and methylation etc.
 Forming disulfide bonds between cysteine residues
 Changes the function

Significance

Proteins are synthesized by ribosomes translating mRNA into

polypeptide chains, which may then undergo modifications to form
the mature protein product.

Post-translational modifications of proteins, which are not gene-

template based, can regulate the protein functions, by causing
changes in protein activity, their cellular locations and dynamic
interactions with other proteins.

PTMs have significant biological functions which include:

Post translational modification
 Aids in proper protein folding – few lectin molecules called
Post-translational modifications, abbreviated s PTMs are the calnexin binds to glycosylated proteins and assist in its
biological modifications that occur after protein synthesis which folding.
alters protein structure and function. Common PTMs are  Confers stability to the protein- glycosylation can modify the
phosphorylation, Methylation, acetylation, lipidation, hydroxylation stability of the protein by increasing protein half life.
and ubiquitination. Post-translational modifications (PTMs) mainly  It protects the protein against cleavage by proteolytic
occur in the endoplasmic reticulum of the cell but sometimes enzyme by blocking the cleavage sites.
continue in the golgi bodies as well. A few common PTMs are  Protein sorting or translocation- If phosphorylated mannose
enlisted here, residues are present in the protein it always goes to
lysosome.
 It regulates protein activity and function- phosphorylation
of protein is a reversible PTM which activates the protein.
 Acetylation regulates many diverse functions, including DNA
recognition, protein-protein interaction and protein
stability.
 Redox-dependent PTM of proteins is emerging as a key
signaling system conserved through evolution, influences
many aspects of cellular homeostasis.
 PTMs are important components in cell signaling, as for
example when prohormones are converted to hormones.
 It significantly increases the diversity and complexity in the
proteome.