Chain Elongation:
Chain elongation during translation is a highly regulated and dynamic process, occurring in three key
steps:
codon recognition,
peptide bond formation
translocation.
Detailed Steps of Chain Elongation:
1. Aminoacyl-tRNA Binding (Codon Recognition)
The first step in elongation involves the delivery of the correct aminoacyl-tRNA (a tRNA molecule
attached to its specific amino acid) to the A site of the ribosome.
Prokaryotes:
o The elongation factor EF-Tu (bound to GTP) guides the aminoacyl-tRNA to the ribosome.
o EF-Tu•GTP complex ensures that the correct aminoacyl-tRNA is placed in the A site by
allowing the anticodon of the tRNA to pair with the corresponding mRNA codon. This
process consumes energy from GTP hydrolysis.
o If the codon-anticodon match is correct, EF-Tu hydrolyzes GTP to GDP and releases the
tRNA.
Eukaryotes:
o The process is similar, but here, the elongation factor eEF1A (bound to GTP) performs
the same role as EF-Tu in prokaryotes.
Regeneration of Elongation Factors:
o After GTP is hydrolyzed to GDP, the elongation factors must be regenerated.
o In prokaryotes, EF-Ts helps regenerate EF-Tu•GTP by exchanging the GDP for a new
GTP, allowing EF-Tu to bind a new aminoacyl-tRNA.
o In eukaryotes, the factor eEF1B regenerates eEF1A•GTP by catalyzing GDP-GTP
exchange.
2. Peptide Bond Formation
Once the aminoacyl-tRNA is correctly placed in the A site, the ribosome catalyzes the formation
of a peptide bond between the amino group of the incoming amino acid in the A site and the
carboxyl end of the polypeptide attached to the tRNA in the P site.
The ribosome's peptidyl transferase activity, which is actually a function of the rRNA in the large
ribosomal subunit (23S rRNA in prokaryotes and 28S rRNA in eukaryotes), catalyzes this
reaction.
After the peptide bond is formed, the growing polypeptide chain is transferred to the tRNA in
the A site.
3. Translocation
In the translocation step, the ribosome moves one codon down the mRNA strand to position the
next codon into the A site.
This causes the following shifts in the ribosomal binding sites:
o The tRNA that was in the P site (now lacking an amino acid) is moved to the E site (exit
site) and then released from the ribosome.
o The tRNA that was in the A site, now carrying the growing polypeptide chain, is shifted
into the P site.
Prokaryotes:
o The elongation factor EF-G (bound to GTP) binds to the ribosome and helps in
translocation by hydrolyzing GTP, providing the energy needed to move the ribosome
one codon forward.
Eukaryotes:
o The corresponding elongation factor is eEF2, which also binds GTP and facilitates
translocation in the same way as EF-G in prokaryotes.
Regeneration of Elongation Factors:
o After translocation, EF-G•GDP (in prokaryotes) or eEF2•GDP (in eukaryotes) must
exchange GDP for GTP to prepare for the next round of translocation. This exchange
does not require an external factor like EF-Ts in the previous step.
Repeating the Cycle
Once the ribosome has moved to the next codon, the A site is now open to accept the next
aminoacyl-tRNA. The cycle continues with the binding of the next aminoacyl-tRNA, the
formation of the next peptide bond, and translocation.
Recap of Key Elongation Factors and Their Roles:
Prokaryotic Eukaryotic
Step Elongation Elongation Function Regeneration
Factors Factors
Brings aminoacyl-tRNA EF-Ts (exchanges GDP for
Aminoacyl-
EF-Tu•GTP eEF1A•GTP to GTP) in prokaryotes;
tRNA Binding
A site eEF1B in eukaryotes
Ribosome (no Ribosome (no Catalyzed by peptidyl
Peptide Bond
elongation elongation transferase (part of large N/A
Formation
factors) factors) ribosomal subunit)
Moves ribosome one
Direct GDP-GTP exchange
Translocation EF-G•GTP eEF2•GTP codon forward
(no factor needed)
(translocation)
Energy Requirement:
Each elongation cycle consumes two GTP molecules:
1. One GTP for the binding of the aminoacyl-tRNA to the A site (via EF-Tu/eEF1A).
2. One GTP for translocation (via EF-G/eEF2).
This ensures that the elongation process is both accurate and efficient, proceeding codon by codon
along the mRNA until the ribosome reaches a stop codon, signaling the end of translation.
Elongation factors and their regeneration
Elongation factors are essential proteins in the elongation phase of translation that facilitate the
accurate and efficient synthesis of proteins. They help in the correct positioning of aminoacyl-tRNAs,
catalysis of peptide bond formation, and translocation of the ribosome along the mRNA. The key
elongation factors and their regeneration in prokaryotes and eukaryotes are:
1. Elongation Factors in Prokaryotes
a. EF-Tu (Elongation Factor Thermo Unstable)
Function: EF-Tu is responsible for delivering the aminoacyl-tRNA (charged tRNA) to the
ribosome's A site. It forms a complex with GTP and the aminoacyl-tRNA, protecting the ester
bond between the amino acid and tRNA.
Mechanism:
1. EF-Tu forms a complex with GTP and the aminoacyl-tRNA.
2. It brings the aminoacyl-tRNA to the A site of the ribosome.
3. If the codon-anticodon match is correct, GTP is hydrolyzed to GDP, and EF-Tu releases
the aminoacyl-tRNA.
Regeneration:
o After hydrolyzing GTP to GDP, EF-Tu is inactivated. To be reused, it needs to exchange
the GDP for a new GTP.
o This regeneration is facilitated by another factor called EF-Ts.
o EF-Ts displaces GDP from EF-Tu, allowing a fresh GTP to bind, recharging EF-Tu for the
next round of aminoacyl-tRNA delivery.
b. EF-G (Elongation Factor G)
Function: EF-G is involved in the translocation of the ribosome. After a peptide bond is formed,
EF-G binds to the ribosome and moves the tRNAs and mRNA down by one codon.
Mechanism:
1. EF-G binds to the ribosome with GTP.
2. GTP hydrolysis provides the energy required for the ribosome to move one codon
forward on the mRNA, shifting the peptidyl-tRNA from the A site to the P site, and the
deacylated tRNA from the P site to the E site.
Regeneration:
o After translocation, EF-G is bound to GDP and must exchange GDP for GTP to be active
again.
o Unlike EF-Tu, EF-G regenerates without the help of an external exchange factor like EF-
Ts. It can directly bind GTP after releasing GDP.
2. Elongation Factors in Eukaryotes
a. eEF1A (Eukaryotic Elongation Factor 1A)
Function: eEF1A performs the same role as EF-Tu in prokaryotes. It delivers aminoacyl-tRNA to
the A site of the ribosome and ensures the correct codon-anticodon pairing.
Mechanism:
1. eEF1A forms a complex with GTP and the aminoacyl-tRNA.
2. It delivers the aminoacyl-tRNA to the A site.
3. If the match is correct, GTP is hydrolyzed to GDP, and eEF1A releases the tRNA, allowing
it to participate in peptide bond formation.
Regeneration:
o Similar to EF-Tu, after GTP hydrolysis, eEF1A must exchange GDP for GTP to be active
again.
o The regeneration of eEF1A is facilitated by another factor, eEF1B, which acts as a
guanine nucleotide exchange factor (GEF). eEF1B promotes the release of GDP from
eEF1A, allowing a new GTP molecule to bind.
b. eEF2 (Eukaryotic Elongation Factor 2)
Function: eEF2 is analogous to EF-G in prokaryotes and is responsible for the translocation step.
It moves the ribosome along the mRNA by one codon after peptide bond formation.
Mechanism:
1. eEF2 binds to the ribosome with GTP.
2. GTP hydrolysis powers the translocation of the ribosome, moving the peptidyl-tRNA
from the A site to the P site and shifting the deacylated tRNA from the P site to the E
site.
Regeneration:
o After GTP is hydrolyzed to GDP, eEF2 must exchange GDP for GTP to become active
again.
o This GDP-GTP exchange for eEF2 happens automatically, without the need for any
additional exchange factors, similar to EF-G in prokaryotes.
Summary of Elongation Factors and Their Regeneration:
Function Prokaryotes Eukaryotes
EF-Tu•GTP delivers aminoacyl- eEF1A•GTP delivers aminoacyl-
Aminoacyl-tRNA delivery
tRNA to A site tRNA to A site
Regeneration of Aminoacyl-tRNA EF-Ts regenerates EF-Tu by eEF1B regenerates eEF1A by
Delivery Factor exchanging GDP for GTP exchanging GDP for GTP
EF-G•GTP moves the ribosome eEF2•GTP moves the ribosome
Translocation of Ribosome
along the mRNA along the mRNA
Regeneration of Translocation EF-G exchanges GDP for GTP eEF2 exchanges GDP for GTP
Factor automatically automatically
Energy Requirements:
Each elongation cycle requires two GTP molecules:
1. One GTP for the binding of the aminoacyl-tRNA to the ribosome (via EF-Tu/eEF1A).
2. One GTP for the translocation of the ribosome (via EF-G/eEF2).