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NUSRAT M G
MSc BIOINFORMATICS
Contents
Animations of Translation
Introduction
Translations in Prokaryotes
Translation in Eukaryotes
MCQs
References
Introduction
Translation:The biosynthesis of a protein or a
polypeptide inside a living cell.
In process of translation the language of nucleotides
sequence on mRNA is translated in to the languageof
amino acid sequence.
It occur in cytoplasm where ribosomes arelocated
It is a universal process
In translation ,messenger RNA is decoded to produce
a specific polypeptide
This uses mRNA sequence as a template to guide the
synthesis of a chain of amino acid that form protein
Many types of transcribed RNA, such
tRNA,rRNA,snRNA are not necessarily translated to
amino acid sequence
Steps of Translation
Translation proceeds in four phases
- Activation
- Initiation
- Elongation and
- Termination
Most studies do not include activation as a step of
translation
Activation
In activation, the correct amino acid iscovalently bonded to the correct tRNA
While technically this not a step in translation, it isrequired for translation to proceed
The amino acid is joined by its carboxyl group to the 3`OH of tRNA by anester bond
with help of ATP
When tRNA has an amino acid linked to it, it istermed as “charged”
Initiation
Prokaryotes initiation require the large and small
subunits, the mRNA, the initiator tRNA and three
initiation factors (IF-1, IF-2, IF-3) and GTP.
IF-3 binds to the free 30s subunit, this help to prevent large
subunit binding to it without mRNA and forming an
inactive ribosome
IF-2 complexed with GTP and IF-1 binds to small subunit .
It will assist the charged initiator tRNA to bind
The 30ssubunit attached to a mRNA molecule making
use of the ribosomal binding site on mRNA
The initiator tRNA can then bind to the complex by base pairing of its
anticodon with AUG codon on mRNA
At this point,IF3 can be released, as its role in keeping the subunits apart are
complete.
This complex is called 30sinitiation complex.
The 50s subunit can now bind, which displace IF1 and IF2, and the GTP is
hydrolysed in this energy consuming step.
This complex is called 70sinitiation complex.
The assembled ribosome has two tRNA bindingsites
These arecalled the A and P sites , for aminoacyl and peptidyl
sites and one site is E(exit site) for empty tRNA
The A siteis where incoming amino acyl tRNA molecule bind,
and the p site is where thegrowing polypeptide chain is usuallyfound
The sites are in cleft of small subunit andcontain adjacent codon that are being
translated.
One major of initiation is the placement of initiator tRNA in the Psite
It is the only tRNA that does this , as all othermust enter the Asite
Elongation
With the formation of 70s initiation complexthe elongation cycle canbegin
Itinvolves three elongation factors EF-Tu,EF-Tsand EF-G,GTP,charged tRNA and the 70sinitiation
complex.
Elongation is divided into 3 steps
1. Amino acyl tRNA delivery.
• EF-TU isrequired to deliver the aminoacyl tRNAto A site and energy is consumed in this step by
hydrolysisofGTP
• The released EF-TuGDP complex is regenerated with the help of EF-TS
• Inthe EF-TuEF-Tsexchange cycle EF-Tsdisplaces the GDP and replace itself by GTP
• The resultant EF-Tu.GTPcomplex is now available tobind another amino acyl tRNA and deliver it
to ribosome.
• All amino acyl tRNA can form this complex withEF-Tuexceptthe initiatortRNA
2 .Peptide bond formation.
• After aminoacyl-tRNA delivery ,the A and P sites are both
occupied and the two amino acids that are to be joined are
close to eachother
• The peptidyl transferase activity of the 50ssubunit can now
form a peptide bond between the two aminoacids
• 3. Translocation
• A complex of EF-G(translocase) and GTP binds to the ribosome
and ,is an energy consuming step, the discharged tRNA is
ejected from the P site, the peptidyl- tRNA is moved from A site
to Psite
• The mRNA moves by one codon relative to one codon to the
ribosome
• GDP and EF-G are released . A new codon is now present in
the vacantsite
Termination
Termination of translation happens when the A site of the ribosome faces a stop codon (UUA,UGA or
UGA)
When this happens , no tRNA can recognize it, but areleasing factor can recognize the stop codons
and causes the release of polypeptidechain
In prokaryotes once a stop codon occupies the A site, three termination or release factor
(RF1,RF2,RF3) contribute tothe hydrolysis of peptidyl-tRNA bond
Releasethe free polypeptide and last unchargedtRNA from P site
The dissociation of the 70s into 30sand 50s subunits
RF1binds A site and release the polypeptide anduncharged tRNA
RF2releases the RF1from A site and release itself aswell from translation binding site(present on
large subunit)
RF3functionunknown
Another factor called Ribosomalreleasingfactor causes the dissociation of 70scomplex
Translation in Eukaryotes
In prokaryotic cell, transcription and translation are
coupled, that is, translation begins while the mRNA is still
being synthesized. In a eukaryotic cell, transcriptionoccur
in the nucleus , and translation occur in thecytoplasm.
Translation process in eukaryotes involve
- Activation ( not essentially the step of translation. This
occur the same way as inprokaryotes)
- Initiation
- Elongation and
- termination
1. Initiation
The initiation of translation in eukaryotes is complex,
involving at least 10initiation factors (eIFs) and is divided
into three steps :
a) Formation of 43spreinitiation complex.
b) Formation of 48s initiation complex.
c) Formation of 80sinitiation complex.
a. Formation of 43s preinitiation complex
• A ternary complex containing met-tRNA and eIF-2 bound to GTP attaches to
40sribosomal subunitto form 43spreinitiation complex.
• The presence of eIF-3 and eIF-1Astabilizes this complex.
b. Formation of 48s initiation complex
• The binding of mRNA to 43s pre-initiation complex results in formation of
48sinitiation complex.
• eIF-4f is formed by the association of eIF-4G,eIF-4A with eIF-4E
• The eIF-4F(referred to as cap binding protein ) binds to the cap ofmRNA.
• Then eIF-4A and eIF-B binds to mRNA and reduces its complexstructure.
• This mRNA is then transferred to 43scomplex
• The ribosomal initiation complex scans the mRNA for identification of
appropriate initiation codon
• 5’-AUG is the initiation codon
c.Formation of 80s initiation complex
• 48sinitiation complex binds to 60sribosomal subunit to form 80sinitiation
complex
• The binding involves hydrolysis of GTP(bound to eIF-2)
• This step is facilitated by the involvement ofeIF-5
• As the 80scomplex is formed, the initiation factors bound to 43sinitiation
complex are released and recycled
2. Elongation
• Ribosomes elongate the polypeptide chain by
sequential addition of aminoacids
• The amino acid sequence is determined by the order of the
codons in the specific mRNA
• Elongation, a cyclic process involving certain
elongation factors(EFs)
• Elongation may be divided into three steps
a. Binding of aminoacyl-tRNA to A-site
b. Peptide bond formation
c. translocation
a. Binding of Aminoacyl t-RNA to A- site
• The 80s initiation complex contains met tRNA in the P-site
and A- site is free
• Another aminoacyl-tRNA is placed in the Asite
• This require proper codon recognition on mRNA and
involvement of EF-1aand supply of energy by GTP
• The aminoacyl –tRNA is placed in the A-site ,EF-1a and
GDP are recycled to bring another aminoacyl- tRNA
b. Peptide bond formation
• The peptidyl transferase catalyzes the formation of
peptide bond
• Net result of peptide bond formation is the attachment off
the growing peptide chain to thetRNA in A-site
c. Translocation
• The ribosome moves to the next codon of mRNA (towards 3’end)
• This process is called translocation, involves the movement of growing
peptide chain from A-site toP- site
• Translocation require EF-2and GTP
• GTP get hydrolyzed and supplies energy to move mRNA
• EF-2and GTP complex recycles for translocation
3. Termination
• One of the stop signals (UGA,UAA and UGA) terminates the growing
polypeptide
• When the ribosome encounter s a stop codon, there isno tRNA available to bind
to the A site of ribosome
• Instead a release factor binds toit
• In eukaryotes eRF1recognizes all the three stop codons, and eRF3stimulate
the termination events.
• Once the release factor binds, the ribosome unit falls apart
• releasing the large and small subunits.
• the tRNA carrying the polypeptide is alsoreleased, freeing up the
polypeptide product.
References
• www.namrata.com (biochemistry For Medics ByDr.
Namrata Chhabra)
• slideshares
• www.easybiologyclass.com
• YouTube lecture by shomou’s Biology
• Wikipedia
Questions/suggestions:
mnusratgulbarga@gmail.com
Thank
You

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Translation in Pro and Eu karyotes

  • 1. NUSRAT M G MSc BIOINFORMATICS
  • 2. Contents Animations of Translation Introduction Translations in Prokaryotes Translation in Eukaryotes MCQs References
  • 3. Introduction Translation:The biosynthesis of a protein or a polypeptide inside a living cell. In process of translation the language of nucleotides sequence on mRNA is translated in to the languageof amino acid sequence. It occur in cytoplasm where ribosomes arelocated It is a universal process
  • 4. In translation ,messenger RNA is decoded to produce a specific polypeptide This uses mRNA sequence as a template to guide the synthesis of a chain of amino acid that form protein Many types of transcribed RNA, such tRNA,rRNA,snRNA are not necessarily translated to amino acid sequence
  • 5. Steps of Translation Translation proceeds in four phases - Activation - Initiation - Elongation and - Termination Most studies do not include activation as a step of translation
  • 6. Activation In activation, the correct amino acid iscovalently bonded to the correct tRNA While technically this not a step in translation, it isrequired for translation to proceed The amino acid is joined by its carboxyl group to the 3`OH of tRNA by anester bond with help of ATP When tRNA has an amino acid linked to it, it istermed as “charged”
  • 7. Initiation Prokaryotes initiation require the large and small subunits, the mRNA, the initiator tRNA and three initiation factors (IF-1, IF-2, IF-3) and GTP. IF-3 binds to the free 30s subunit, this help to prevent large subunit binding to it without mRNA and forming an inactive ribosome IF-2 complexed with GTP and IF-1 binds to small subunit . It will assist the charged initiator tRNA to bind The 30ssubunit attached to a mRNA molecule making use of the ribosomal binding site on mRNA
  • 8. The initiator tRNA can then bind to the complex by base pairing of its anticodon with AUG codon on mRNA At this point,IF3 can be released, as its role in keeping the subunits apart are complete. This complex is called 30sinitiation complex. The 50s subunit can now bind, which displace IF1 and IF2, and the GTP is hydrolysed in this energy consuming step. This complex is called 70sinitiation complex. The assembled ribosome has two tRNA bindingsites These arecalled the A and P sites , for aminoacyl and peptidyl sites and one site is E(exit site) for empty tRNA The A siteis where incoming amino acyl tRNA molecule bind, and the p site is where thegrowing polypeptide chain is usuallyfound
  • 9. The sites are in cleft of small subunit andcontain adjacent codon that are being translated. One major of initiation is the placement of initiator tRNA in the Psite It is the only tRNA that does this , as all othermust enter the Asite
  • 10. Elongation With the formation of 70s initiation complexthe elongation cycle canbegin Itinvolves three elongation factors EF-Tu,EF-Tsand EF-G,GTP,charged tRNA and the 70sinitiation complex. Elongation is divided into 3 steps 1. Amino acyl tRNA delivery. • EF-TU isrequired to deliver the aminoacyl tRNAto A site and energy is consumed in this step by hydrolysisofGTP • The released EF-TuGDP complex is regenerated with the help of EF-TS • Inthe EF-TuEF-Tsexchange cycle EF-Tsdisplaces the GDP and replace itself by GTP • The resultant EF-Tu.GTPcomplex is now available tobind another amino acyl tRNA and deliver it to ribosome. • All amino acyl tRNA can form this complex withEF-Tuexceptthe initiatortRNA
  • 11. 2 .Peptide bond formation. • After aminoacyl-tRNA delivery ,the A and P sites are both occupied and the two amino acids that are to be joined are close to eachother • The peptidyl transferase activity of the 50ssubunit can now form a peptide bond between the two aminoacids • 3. Translocation • A complex of EF-G(translocase) and GTP binds to the ribosome and ,is an energy consuming step, the discharged tRNA is ejected from the P site, the peptidyl- tRNA is moved from A site to Psite • The mRNA moves by one codon relative to one codon to the ribosome • GDP and EF-G are released . A new codon is now present in the vacantsite
  • 12.
  • 13. Termination Termination of translation happens when the A site of the ribosome faces a stop codon (UUA,UGA or UGA) When this happens , no tRNA can recognize it, but areleasing factor can recognize the stop codons and causes the release of polypeptidechain In prokaryotes once a stop codon occupies the A site, three termination or release factor (RF1,RF2,RF3) contribute tothe hydrolysis of peptidyl-tRNA bond Releasethe free polypeptide and last unchargedtRNA from P site The dissociation of the 70s into 30sand 50s subunits RF1binds A site and release the polypeptide anduncharged tRNA RF2releases the RF1from A site and release itself aswell from translation binding site(present on large subunit) RF3functionunknown Another factor called Ribosomalreleasingfactor causes the dissociation of 70scomplex
  • 14.
  • 15. Translation in Eukaryotes In prokaryotic cell, transcription and translation are coupled, that is, translation begins while the mRNA is still being synthesized. In a eukaryotic cell, transcriptionoccur in the nucleus , and translation occur in thecytoplasm. Translation process in eukaryotes involve - Activation ( not essentially the step of translation. This occur the same way as inprokaryotes) - Initiation - Elongation and - termination
  • 16. 1. Initiation The initiation of translation in eukaryotes is complex, involving at least 10initiation factors (eIFs) and is divided into three steps : a) Formation of 43spreinitiation complex. b) Formation of 48s initiation complex. c) Formation of 80sinitiation complex.
  • 17. a. Formation of 43s preinitiation complex • A ternary complex containing met-tRNA and eIF-2 bound to GTP attaches to 40sribosomal subunitto form 43spreinitiation complex. • The presence of eIF-3 and eIF-1Astabilizes this complex. b. Formation of 48s initiation complex • The binding of mRNA to 43s pre-initiation complex results in formation of 48sinitiation complex. • eIF-4f is formed by the association of eIF-4G,eIF-4A with eIF-4E • The eIF-4F(referred to as cap binding protein ) binds to the cap ofmRNA. • Then eIF-4A and eIF-B binds to mRNA and reduces its complexstructure. • This mRNA is then transferred to 43scomplex • The ribosomal initiation complex scans the mRNA for identification of appropriate initiation codon • 5’-AUG is the initiation codon
  • 18. c.Formation of 80s initiation complex • 48sinitiation complex binds to 60sribosomal subunit to form 80sinitiation complex • The binding involves hydrolysis of GTP(bound to eIF-2) • This step is facilitated by the involvement ofeIF-5 • As the 80scomplex is formed, the initiation factors bound to 43sinitiation complex are released and recycled
  • 19. 2. Elongation • Ribosomes elongate the polypeptide chain by sequential addition of aminoacids • The amino acid sequence is determined by the order of the codons in the specific mRNA • Elongation, a cyclic process involving certain elongation factors(EFs) • Elongation may be divided into three steps a. Binding of aminoacyl-tRNA to A-site b. Peptide bond formation c. translocation
  • 20. a. Binding of Aminoacyl t-RNA to A- site • The 80s initiation complex contains met tRNA in the P-site and A- site is free • Another aminoacyl-tRNA is placed in the Asite • This require proper codon recognition on mRNA and involvement of EF-1aand supply of energy by GTP • The aminoacyl –tRNA is placed in the A-site ,EF-1a and GDP are recycled to bring another aminoacyl- tRNA
  • 21. b. Peptide bond formation • The peptidyl transferase catalyzes the formation of peptide bond • Net result of peptide bond formation is the attachment off the growing peptide chain to thetRNA in A-site c. Translocation • The ribosome moves to the next codon of mRNA (towards 3’end) • This process is called translocation, involves the movement of growing peptide chain from A-site toP- site • Translocation require EF-2and GTP • GTP get hydrolyzed and supplies energy to move mRNA • EF-2and GTP complex recycles for translocation
  • 22.
  • 23. 3. Termination • One of the stop signals (UGA,UAA and UGA) terminates the growing polypeptide • When the ribosome encounter s a stop codon, there isno tRNA available to bind to the A site of ribosome • Instead a release factor binds toit • In eukaryotes eRF1recognizes all the three stop codons, and eRF3stimulate the termination events. • Once the release factor binds, the ribosome unit falls apart • releasing the large and small subunits. • the tRNA carrying the polypeptide is alsoreleased, freeing up the polypeptide product.
  • 24.
  • 25. References • www.namrata.com (biochemistry For Medics ByDr. Namrata Chhabra) • slideshares • www.easybiologyclass.com • YouTube lecture by shomou’s Biology • Wikipedia Questions/suggestions: mnusratgulbarga@gmail.com