DNA Translation Flashcards
Describe the genetic code.
They are redundant (more than 1 codon can code for same amino acid), non-overlapping, unambigious and universal.
64 codons, 61 code for amino acids and 3 triplets for stop codons in translation.
They must be read in the correct reading frame.
Describe the structure of tRNA.
A single RNA strand abount 80 nucleotides long.
When flattened into 1 plane, tRNA looks like a cloverleaf. Although its L-shaped in 3D.
Each has a specific amino acid at one end and an anticodon at the other, this pairs with complementary codons on mRNA.
What are the 2 events needed for accurate translation?
Correct match between tRNA and amino acid, this is done by enzyme aminoacyl-tRNA synthetase.
Correct match between tRNA anticodon and mRNA codon.
What is the ‘wobble’?
The third base of codon is flexible in pairing with anticodon, so some tRNAs can bind with more than 1 codons. This flexible pairing is called wobble.
How are tRNAs synthesised and modified?
tRNAs transcribed by RNA polymerase III as precursor molecules that need to be spliced and modified.
They are covalently modified before exit from the nucleus (1/10 nt is modified in mature tRNAs; more than 50 types of modifications).
Describe the binding sites of ribosomes.
A (amino acid) site - holds tRNA that carries the next amino acid to be added.
P (polypeptide) site - holds tRNA that carries the growing nucleotide chain.
E (exit) site - discharged tRNAs leave the ribosome.
How is protein synthesis initiated?
A small ribosomal subunit binds to the mRNA and an initiator tRNA.
Small subunit moves along mRNA until it reaches a start codon (AUG).
Initiation factos bring a large ribosomal unit to complete translation initiation complex.
How can bacteria translate multiple peptides from a single mRNA?
In prokaryotic cells there is usually a polycistronic messenger. This means that more than 1 proteins are encoded by a singular mRNA.
Consequently they have more than 1 ribosome binding sites surrounding specific initiator codons for each specific polypeptide chain to be translated.
What happens during the elongation stage?
Amino acids are added one by one to the preceding amino acid.
Each addition involves proteins called elongation factors and occurs in 3 steps: codon recognition, peptide bond formation, and translocation.
Describe the formation of the initiation complex.
Initiator tRNA binds with methionyl and a small ribosomal subunit. There is also an initiation factor elF2 bound to GTP.
Initiation factor tRNA is bound to the P site (not usually the case). This is the pre-complex.
The pre-comlpex shifts along the mRNA (circular structure) using ATP to reach the start codon (AUG). There are more than 1 AUG, so they are skipped until the correct one is determined (embedded in a correct context, or sequence around it).
The GTP bound to elF2 will be hydrolyzed to start initiation.
Large ribosomal subunit binds to form the initiation complex.
The 2nd tRNA (aminoacyl-tRNA) will arrive with its amino acid at the A site. This amino acid is bound to methionyl via a peptide bond.
Describe the general process of the elongation stage.
The amino acid of the newly bound charged tRNA will need to make a peptide bond with the previous amino acid in the polypeptide chain.
The entire chain will need to shift over with the now bound polypeptide tRNA occupying the P-site, and the previous tRNA will be released from the E-site.
Then a newly bound charged tRNA will repeat the process and grow the polypeptide chain to eventually reach a stop codon.
Specific elongation factors needed for this process.
Describe the role of the elongation factors in elongation.
EF-Tu (Elongation factor Thermo Unstable):
- Bound to GTP, therefore has high affinity for aa-tRNA.
- Forms the ternary complex.
- It brings the loaded tRNA into the A-site.
- Can also proofread by checking if hydrolyzation between anticodon and codon is correct. If not, it can aid in their dissociation.
EF-G:
- Translocates the tRNA and mRNA complex.
- Moves the whole mRNA-tRNA complex along by 1 codon. I.e. The peptidyl-tRNA from A to P and deacetylated tRNA from P to E.
Describe the structure of the ribosome.
The ribosome is a ribozyme. It’s made of proteins and RNAs.
RNA is the catalytic part of the ribosome.
16S (18S in eukaryotic cells) rRNA brings together codons and anticodons and the 23S rRNA catalyzes formation of peptide bonds.
Describe the process of termination.
Stop codons mark the end of translation.
Occurs when a stop codon reaches the A site
A site accepts protein called a release factor.
Release factor leads to the addition of a water molecule instead of an amino acid.
This reaction releases the polypeptide and the translation assembly comes apart.
Describe the process of termination.
Stop codons mark the end of translation.
Occurs when a stop codon reaches the A site, then A site accepts the protein called release factor.
Release factor leads to the addition of a water molecule instead of an amino acid.
This reaction releases the polypeptide and the translation assembly comes apart.