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Translation
production of a protein from the info in an mRNA
Transfer RNA (tRNA)
read the mRNA by base-pairing anticodon with codon; translate mRNA into protein; carry specific aa at the 3' end
Aminoacyl-tRNA synthetase
Attach aas to tRNAs
Ribosome
carry out translation; 1/3 protein and 2/3 RNA; move 5'-3' along mRNA
Small subunit of ribosome
Deciphers mRNA
Large subunit of ribosome
Mediates chemical bond formations
Translation factors
Often called GTPases; associate with ribosomes and help with translation
4 stages of translation
Initiation, elongation, termination, ribosome recycling
tRNA structure
4 regions of dsRNA with 3 stem-loops
Acceptor stem
5' and 3' ends of tRNA base-pair; conserved 3' CCA tail that binds the aa
Anticodon loop
3 nucleotides that base pair with the 3 nucleotides of the codon in mRNA
Triplet code
Each codon specifies a single aa (sense) or no aa (stop/ nonsense); simplest code that can specify all 20 aas; some aas encoded by more than one codon
Isoacceptors
Different tRNAs that carry the same aa
Reading the 3 mRNA positions
First 2 are read by strict Watson-Crick base pairing with positions 2 and 3 of the anticodon; 3rd interacts with position 1 of the anticodon via wobble pairing
Genetic code
Almost the same in all organisms (AUG-methionine; UAA, UAG, and UGA-stop); mutations that change the aa encoded usually result in a similar aa taking its place
Aminoacylation process
2 steps requiring ATP that attach aas to tRNAa; fewer than 1 error per 104 aminoacylation events
Identity elements
sequence and structural features by which aminoacyl-tRNA synthetases identify tRNAs
Exit tunnel
part of large subunit where growing polypeptide emerges; often a target for antibiotics
Interface between subunits
important for movement of tRNAs and mRnA
3 tRNA binding sites in ribosome
aminoacyl (A) site, peptidyl (P) site, exit (E) site
Initiation
AUG identified by initiation factors (IFs), ribosome, and special initiator methionine tRNA; methionine-loaded tRNA is bound to P site
Elongation
Elongation factor Tu (eEF1A) loads next charged tRNA into A site; peptide bond formation catalyzed between aa in P and aa in A; EFG (EF2) then promotes translocation
Translocation
Movement of mRNA-tRNA through ribosome; peptidyl-tRNA in A site moves to P site, new codon moves to A site, tRNA in E site leaves ribosome
Termination
ribosome reaches a stop codon (UAG, UAA, or UGA); stop codon recognized by release factors; promotes release of polypeptide from ribosome
Class I release factors that recognize stop codons
In bacteria, RF1 for UAA and UGA, RF2 for UAA and UGA In eukaryotes, eRF1 for all 3
Ribosome recycling
Large and small subunits dissociate and release remaining tRNA and mRNA; recycling factor RRF and EFG help dissociation in bacteria
3 steps of translation initiation
small subunit identifies start codon in mRNA; methionyl-tRNA is loaded into P site and base pairs with start codon; large subunit joins complex
Initiator tRNA in eukaryotes vs. bacteria
decodes AUG initiator codon; tRNA1Met in eukaryotes and tRNAfMet in bacteria, f denoting a formyl group
Polycistronic
Bacterial mRNAs have several open reading frames, each with its own start and stop codon
Shine-Dalgarno sequence
Polypurine tract 6-8 bases upstream of initiator AUG in bacteria; pairs with polypyrimidine region in 3' end of 16S rRNA (anti-Shine-Dalgarno sequence); guides initiator AUG to P site
Shine-Dalgarno Consensus
AGGAGGU; sequence deviations from consensus control strength of translation
Differences between eukaryotic initiation and bacterial initiation
No eukaryotic equivalents to Shine-Dalgarno sequence; small subunit does not bind directly to mRNA; more initiation factors needed; monocistronic mRNAs; initiation can occur at 2nd and 3rd AUG in addition to 1st
3 steps of elongation
Decoding, peptide bond formation, translocation
Decoding
aminoacyl-tRNA with anti-codon complementary to mRNA codon is chosen by the ribosome
Peptide bond formation
transfer of polypeptide chain to the aminioacyl-tRNA in the A site
Ribosome recycling factor (RRF)
In bacteria, promotes ribosome disassembly
Nonsense suppression
stop codons are misread and termination fails to occur
Frameshifting
mRNA shifts so that peptide synthesis proceeds in a different reading frame
Antibiotics
Small molecules that kill or disrupt organism growth; most effective target bacterial or fungal process but don't disrupt the same process in mammals
Antibiotics and translation
Small differences between bacterial and eukaryotic translation allow good antibiotics to target the ribosome and other translation proteins

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