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Lecture 19:
Translation in eukaryotes
Readings (chapter 6)
Course 281
Lessons for life
AIMS
• Understand the process of translation in
eukaryotes.
• Understand the molecular requirements to
translate a eukaryotic mRNA into a protein.
• Understand the sequence of events in eukaryotic
translation.
• Understand the differences between prokaryotic
and eukaryotic translation.
Summary and review
Prokaryotes Eukaryotes
No nucleus Nucleus
DNA in cytoplasm DNA in nucleus
Transcription in cytoplasm Transcription in nucleus
Translation in cytoplasm Translation in cytoplasm
Polcistronic transcripts
(one transcript many genes)
Monocistronic transcript
(one transcript one gene)
Coupled transcription and
translation
Transcription and translation
NOT coupled
mRNA not processed RNA processed
One RNA polymerase Many RNA polymerases
Summary and review
Review
Eukaryotic mRNA
Eukaryotic translation
• Eukaryotic translation is similar to that of
prokaryotes but a bit more complex.
• Differences in the translation process is due
to differences in transcript structure and
location of translation.
Eukaryotic translation
What are the stages of protein synthesis in
eukaryotes?
(1) Initiation
(2) Elongation
(3) Termination
Eukaryotic translation initiation
What is needed for translation initiation?
1.Mature mRNA
2.Ribosome
3.Eukaryotic initiation factors (eIFs)
4.GTP
5.Initiator tRNA (Met-tRNA)
Eukaryotic translation initiation
Differences between eukaryotic and
prokaryotic translation initiation?
1.No Shine-Dalgarno sequence in eukaryotes
2.No modified initiator tRNA (fMet-tRNA)
How does the ribosome know where to start?
Eukaryotic translation initiation
The process:
1.Initiator tRNA (Met-tRNA) gets loaded onto the
small ribosomal subunit (40S).
2.Eukaryotic initiation factors (eIFs) gets loaded as
well on the 40S ribosomal subunit.
3.A pre-initiation complex is formed containing
(40S – Met-tRNA – eIFs).
Eukaryotic translation initiation
Eukaryotic translation initiation
Note:
Why only Met-tRNA binds to the 40S
ribosomal subunit?
What about other charged tRNAs?
Only Met-tRNA can bind tightly to the small
ribosomal subunit without the assembly of the
entire ribosome.
Eukaryotic translation initiation
The process:
4.The complex (40S – Met-tRNA – eIFs) binds to
the 5’ end of the mRNA (5’ Cap).
5.More eIFs are bound to the 5’ Cap.
Eukaryotic translation initiation
The process:
6.The complex (40S –
Met-tRNA – eIFs) moves
along the transcript (5’ –
3’) in search for the first
AUG.
7.The process is called
“scanning” of mRNA.
8.When first AUG is found,
eIFs dissociate.
Eukaryotic translation initiation
The process:
9.The large ribosomal subunit (60S) assembles to
the complex at the start codon.
10. The initiator tRNA is positioned in the P site of
the ribosome and the A site is available to receive
to next charged tRNA.
Summary of initiation
Summary
Translation initiation in
eukaryotes
Eukaryotic translation elongation
Eukaryotic translation termination
• Termination in
eukaryotes is similar
to that in prokaryote.
• Eukaryotic Release
factor (eRF1)
recognizes all stop
codons.
• Other release factors
help in the termination
and disassembling of
the ribosome.
Coupled transcription translation in bacteria
Polysome
Polysome or polyribosome: a number of
ribosomes translating the same transcript.
Polysome
• Polysome or polyribosome: a number of
ribosomes translating the same transcript.
• Many rounds of translation takes place on the
same transcript.
Why is this process beneficial for the cell?
• Saves time (How long)
• Quantity (How much)
Polycistronic transcript
Translating polycistronic prokaryotic
transcript:
• Each gene gets translated independently
because each has its own Shine-Dalgarno
sequence.
Expectations
• You know how translation process occurs in
eukaryotes.
• You know how ribosome finds the correct
location to start translation in eukaryotes.
• You know the molecules needed in every step of
translation.
• You the sequence of events in eukaryotic
translation.
• You know the differences between prokaryotic
and eukaryotic translation.
For a smile

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281 lec19 euk_translation

  • 1. Lecture 19: Translation in eukaryotes Readings (chapter 6) Course 281
  • 3. AIMS • Understand the process of translation in eukaryotes. • Understand the molecular requirements to translate a eukaryotic mRNA into a protein. • Understand the sequence of events in eukaryotic translation. • Understand the differences between prokaryotic and eukaryotic translation.
  • 4. Summary and review Prokaryotes Eukaryotes No nucleus Nucleus DNA in cytoplasm DNA in nucleus Transcription in cytoplasm Transcription in nucleus Translation in cytoplasm Translation in cytoplasm Polcistronic transcripts (one transcript many genes) Monocistronic transcript (one transcript one gene) Coupled transcription and translation Transcription and translation NOT coupled mRNA not processed RNA processed One RNA polymerase Many RNA polymerases
  • 7. Eukaryotic translation • Eukaryotic translation is similar to that of prokaryotes but a bit more complex. • Differences in the translation process is due to differences in transcript structure and location of translation.
  • 8. Eukaryotic translation What are the stages of protein synthesis in eukaryotes? (1) Initiation (2) Elongation (3) Termination
  • 9. Eukaryotic translation initiation What is needed for translation initiation? 1.Mature mRNA 2.Ribosome 3.Eukaryotic initiation factors (eIFs) 4.GTP 5.Initiator tRNA (Met-tRNA)
  • 10. Eukaryotic translation initiation Differences between eukaryotic and prokaryotic translation initiation? 1.No Shine-Dalgarno sequence in eukaryotes 2.No modified initiator tRNA (fMet-tRNA) How does the ribosome know where to start?
  • 11. Eukaryotic translation initiation The process: 1.Initiator tRNA (Met-tRNA) gets loaded onto the small ribosomal subunit (40S). 2.Eukaryotic initiation factors (eIFs) gets loaded as well on the 40S ribosomal subunit. 3.A pre-initiation complex is formed containing (40S – Met-tRNA – eIFs).
  • 13. Eukaryotic translation initiation Note: Why only Met-tRNA binds to the 40S ribosomal subunit? What about other charged tRNAs? Only Met-tRNA can bind tightly to the small ribosomal subunit without the assembly of the entire ribosome.
  • 14. Eukaryotic translation initiation The process: 4.The complex (40S – Met-tRNA – eIFs) binds to the 5’ end of the mRNA (5’ Cap). 5.More eIFs are bound to the 5’ Cap.
  • 15. Eukaryotic translation initiation The process: 6.The complex (40S – Met-tRNA – eIFs) moves along the transcript (5’ – 3’) in search for the first AUG. 7.The process is called “scanning” of mRNA. 8.When first AUG is found, eIFs dissociate.
  • 16. Eukaryotic translation initiation The process: 9.The large ribosomal subunit (60S) assembles to the complex at the start codon. 10. The initiator tRNA is positioned in the P site of the ribosome and the A site is available to receive to next charged tRNA.
  • 17. Summary of initiation Summary Translation initiation in eukaryotes
  • 19. Eukaryotic translation termination • Termination in eukaryotes is similar to that in prokaryote. • Eukaryotic Release factor (eRF1) recognizes all stop codons. • Other release factors help in the termination and disassembling of the ribosome.
  • 21. Polysome Polysome or polyribosome: a number of ribosomes translating the same transcript.
  • 22. Polysome • Polysome or polyribosome: a number of ribosomes translating the same transcript. • Many rounds of translation takes place on the same transcript. Why is this process beneficial for the cell? • Saves time (How long) • Quantity (How much)
  • 23. Polycistronic transcript Translating polycistronic prokaryotic transcript: • Each gene gets translated independently because each has its own Shine-Dalgarno sequence.
  • 24. Expectations • You know how translation process occurs in eukaryotes. • You know how ribosome finds the correct location to start translation in eukaryotes. • You know the molecules needed in every step of translation. • You the sequence of events in eukaryotic translation. • You know the differences between prokaryotic and eukaryotic translation.