Transcription and Translation Flashcards
What are the 4 types of RNA?
mRNA
rRNA
tRNA
ncRNA
What is the structure and lifetime of mRNA?
Unstructured
Short-lived (minutes)
What is the structure and lifetime of rRNA?
Highly structured
Long-lived
What is the structure and lifetime of tRNA?
Identical structures
Long-lived
What is the structure of tRNA?
They are identical structures
There is an amino acid attachment site
Anti-codon
What is tRNA aminoacylation?
Amino acids are added based on the anticodon
What is rRNA structure?
70s structure to 30s and 50s
30s structure to 16s
50s structure to 23s and 5s
What do most bacterial RNA polymerases have?
A core enzyme composed of 5 chains that catalyze RNA synthesis
What are sigma factors?
Have no catalytic activity but help the core enzyme recognize the start of genes
What is a holoenzyme?
A core enzyme + the sigma factor
Only the holoenzyme can begin transcription
What do sigma factors do?
Help RNA polymerase bind to the promotor
What is the promotor?
The site where RNA polymerase binds to initiate transcription
Are promotors transcribed?
No
They have specific sequences before the transcription starting point and a Pribnow box which contains consensus sequence
What is the transcription bubble?
Moves with the polymerase as it transcribes mRNA from the template strand
Within the bubble a temporary RNA:DNA hybrid is formed
When does termination occur?
When core RNA polymerase dissociates from template DNA
Where do DNA sequences mark the end of the gene?
In the trailer and the terminator
What do some terminators require?
The rho factor for termination
What are you left with after transcription?
Leaves you with free mRNA
Are there 5’ caps or long poly-A tails in bacteria?
No
What is a gene?
The basic unit of genetic information
What is a locus?
Position on a genome
What is a gene cluster?
Cluster of gene encoding proteins involved in the biosynthesis of one structure
What is an operon?
Cotranscribed
One mRNA for all the genes
How do you tell if something is an operon?
Through experimentation
What is a start codon?
Start site for translation
Begins with DNA sequence 3’-TAC-5’
Produces codon AUG
What is a stop codon?
Coding region ends with a stop codon
UAA, UAG, UGA
Do not encode amino acids
What is code degeneracy?
Up to 6 different codons can code for a single amino acid
What is codon usage bias?
There are fewer tRNAs in the tRNA pool for some codons which slows down translations at these sites
What is 3rd base pair wobble?
Loose base pairing in the 3rd position of the codon
Eliminates the need for unique tRNA for each codon
What are reading frames?
Specifies where the codons are
The ribosome binding site is always 6-7 bp upstream of the start of the codon
What is tRNA aminoacylation?
Amino acids are added based on the anticodon
Attached via the carboxyl group
Requires ATP hydrolysis
What are the steps of translation initiation?
- Binding of mRNA to 30S subunit
- Binding of initiator tRNA to complex
- Hydrolysis of GTP by initiation factors to bring 50S subunit
What sites does an intact ribosome contain?
A site (amino acid) P site (peptide) E site (exit)
What is the elongation cycle?
Sequential addition of amino acids to growing peptide
What are the 3 phases of elongation?
Aminoacyl-tRNA binding
Transpeptidation reaction
Translocation
What does the peptidyl (P) site do?
Binds initiator tRNA or tRNA attached to the growing polypeptide
What does the aminoacyl (A) site do?
Binds incoming aminoacyl-tRNA
What does the exit (E) site do?
Briefly binds empty tRNA before it leaves ribosome
What is aminoacyl-tRNA binding?
Aminoacyl-tRNA enters the ribosome
Based on codon
Requires EF-Tu and GTP hydrolysis
What is transpeptidation?
Formation of the peptide bond
Catalyzed by peptidyl transferase activity of 23S rRNA
Amino groups on the A site amino acid performs a nucleophilic attack on the carboxyl group of the C-terminal amino acid on the P site of tRNA
What are the 3 simultaneous events in translocation?
Peptidyl-tRNA moves from A site to P site
The ribosome moves down one codon
Empty tRNA moves to E site and dissociates
What does translocation require?
Requires EF-G (translocase) and GTP hydrolysis
What is termination?
Takes place at any of the 3 nonsense codons
Has release factors
Requires GTP hydrolysis
Ribosome dissociates into 30S and 50S components
What are release factors?
Aid in recognition of stop codons
3 RFs function in prokaryotes
Only 1 RF active in eukaryotes