genetic code
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Genetic code andprotein synthesis
Lodish Chapter 4.4
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Pol I transcriptprocessed bysmall nucleolarribonucleoproteins(snoRNPs)
Pol III transcript
S (Svedberg unit): sedimentation coefficient
Ribosomal RNAis transcribed and processed in thenucleolus
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All aminoacyl-tRNA synthetases have proofreading activity
enzyme tRNA binding highly dependenton tertiary structure of tRNA
if product doesn’t fit correctlythen proofreading activity
reverses bondacceptor stem
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3 letter code allows for 3 different reading frames
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64 possible codons, 61 of which encode for 20 amino acids and 3 for Stops
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Some exceptions to the rule
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The base pair in the 3rd codon position can “wobble”
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60% 40%
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Initiation factors (eIFs) regulate ribosome assembly
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Polyadenylation mostlyin nucleus, but can alsooccur in cytoplasm
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Selection of the initiating AUG is determined by neighboring nucleotides
In eukaryotes: ACCAUGG (Kozak sequence)or internal ribosomal entry sites (IRES-typical in viruses with long5’ untranslated regions)
Shine-Delgarno
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Crystal structure of prokaryotic ribosome reveals threedistinct tRNA binding sites: A, P and E
Switches in rRNAinteractionspromoteconformationalchanges
RNA-drivenmachine
peptide bondformationrequirescontactbetween23S rRNAand tRNAacceptor stem(can be carried outw/o proteins in vitro)
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Elongation factors directtRNAs to the A site
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Release factors recognizeSTOP codons
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frameshift mutations
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What you need to know:Ribosome assembly: rRNAs are processed by snoRNPs in the nucleolusSubunit composition of ribosomes in prokaryotic/ eukaryotic cellsUniversal code:64 codons, 3rd position is wobble position. Definition of codon and anticodon.tRNAs are activated by aminoacyl tRNA synthetases, which have proof-reading activity (only 1 in 50,000 is wrong)First AUG is located at a defined distance to the Shine-Delgarno sequence, which is a ribosomal docking site.Translation initiation:5’cap binds eIF4, which in turn recruits 30S/40S subunit to mRNA. Then first tRNA binds, and then the 50S/60S subunit binds.First tRNA is always tRNA-MET. First AUG is positioned over P site3 tRNA binding sites: A (aminoacyl), P (peptide bond) and E (exit)Elongation requires elongation factors, termination requires releasefactors. Release factors bind to STOP codons.Antibiotics inhibit aminoacyl-tRNA binding, tRNA transposition or cause frameshift mutations and premature chain termination.