Translation Flashcards
Translation
Biosynthesis of proteins based on mRNA
What is needed for translation
- mRNA
- tRNA
- Ribosomes
- Protein factors
Function of: mRNA
Carries genetic information
Function of: tRNA
Transports activated amino acids for translation
What process takes place in the ribosome?
Translation
What forms the structure of tRNA?
Intramolecular base pairing
Give the three loop types found in tRNA
- DHU-loop
- Anticodon-loop
- TφC-loop
DHU-loop binds…
Aminoacyl-tRNA-synthetase
Anticodon-loop binds…
Codons of mRNA
TφC-loop binds
The large subunit of the ribosome
The T in the TφC-loop represents…
Thymidine-monophosphate
Thymine in RNA
The φ in the TφC-loop represents…
Pseudouridine-monophosphate
Describe the binding of a tRNA with an amino acid
- CCA-sequence (3’-end) of tRNA
Binds to
- Carboxyl group of amino acid
How are amino acids activated?
Binding of the adequate amino acid with the tRNA
Name the enzyme which binds the amino acid to its specific tRNA
Aminoacyl-tRNA-synthetase
Aminoacyl-tRNA-synthetase binds…
ATP
Give the reaction of amino acid binding to tRNA
- ATP + Amino acid → Aminoacyl-AMP + PPi
- Aminoacyl-AMP + tRNA → Aminoacyl-tRNA + AMP
Aminoacyl-tRNA-synthetase binds to which part of tRNA?
DHU-loop
Give the structure of a ribosome
- Proteins
- Ribosomal RNA (rRNA)
- Large subunit
- Small subunit
Different types of rRNA can be characterised by their…
- Sedimentation coefficient
- Unit: Svedberg = S
Give the binding sites of large ribosomal subunits
- A-site
- P-site
- E-site
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A-binding site
Aminoacyl-tRNA binding site
P-binding site
Peptidyl-tRNA binding site
E-binding site
Exit
Give the binding site of small ribosomal subunits
mRNA-binding site
In eukaryotes, translation occurs in the…
- Cytoplasm
- RER (membrane with ribosomes)
Summarise the steps of initiation in eukaryotes
- Pre-initiation complex
- Kozak-scanning mechanism
- Recognised start codon
- Initiation complex
Give the composites of the pre-initiation complex
- Small ribosomal subunit
- eIF-2-GTP
- other eIF-S
- tRNAMet
Kozak-scanning mechanism
- Pre-initiation complex rolls along mRNA strand
- Locates the start codon (AUG)
Recognised start codon stage
Small ribosomal subunit binds to Kozak-sequence of mRNA
Give the formation of the initiation complex
- Hydrolysis of the GTP bound to eIF-2 → GDP + Pi
- Large ribosomal subunit binds to the small
- All eIF-s dissociate
What occurs at the end of initiation
- Methionyl-tRNA binds via TφC-loop to P-site of the large subunit
- A-site of the large ribosomal subunit is free
Steps of elongation of translation in eukaryotes
- Binding of aminoacyl-tRNA to A-site of large ribosomal subunit
- Methionine moved from methionyl-tRNA onto the newly bound aminoacyl-tRNA
- Translocation
Describe binding of aminoacyl-tRNA to A-site of the large ribosomal subunit
- EF-1α + GTP complex needed
-
Aminoacyl-tRNA binds to:
- The A-site of large RSU via TφC-loop
- The next codon of mRNA via its anticodon loop
Describe methionine placement onto the aminoacyl-tRNA
- Catalysed by Peptidyl-transferase
- A new peptide bond is formed between:
- Methionine
- Newly bound amino acid
- Forms a dipeptidyl-tRNA
- The ‘empty’ tRNA moves from P-site to the E-site
- It then dissociates
Translocation stage of eukaryotic translation
- Dipeptidyl-tRNA placed onto the P-site
- Catalysed by translocase
- Requires:
- EF-2
- GTP complex
Describe the end stage of eukaryotic translation
- Dipeptidyl-tRNA binds to the P-site of the large RSU via TφC-loop
- A-site of the large ribosomal subunit is free
Describe termination of translation in pro- and eukaryotes
- Stop codon: no aminoacyl-tRNA can bind to A-site
- PRF protein binds to A-site instead
- Deliberation of the newly synthesised protein chain
- By dissociation of ribosomal subunits
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Initiation phase of translation in prokaryotes
- Small RSU + IF-1 and IF-3
- 30S initiation complex formation
- 70S initiation complex
30S initiation complex is formed by
- Small ribosomal subunit + IF-1 and IF-3
- IF-2-GTP + tRNAini + mRNA
How is the 70S initiation complex formed
- IF-1 and IF-3 dissociate
- GTP hydrolyses
- Large RSU binds
List the major differences between the prokaryote and eukaryote stage of initiation
- 3 IF-s are needed for prokaryotes, Eukaryotes need more
- Initiator amino acid: N-formyl-methionine in prokaryotes
- No Kozak-scanning mechanism in prokaryotes
Give the elongation stage of prokaryotic translation
- Different elongation factors:
- EF-Tu; EF-G
- Transcription + translation happen simultaneously
- One mRNA binds to more than one ribosome
- Polyribosome
What is shown in the figure
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Polyribosome
Energy requirement of: Initiation
1 GTP
Energy requirement of: Kozak-scanning mechanism
1 ATP
Energy requirement of: Elongation
4 ATP
Energy requirement of: Termination
1 GTP