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Outline Major biomacromolecules involving the translation General Properties of translation Detailed mechanism of translation Bacterial translation Eukaryotic translation Archaeal translation Quality control of mRNA Translation inhibitors Intracellular proteolysis Functional sites on Ribosome Three sites for tRNA The A (acceptor) site - where aa-tRNAs come in (except the first one) and where peptidyl-tRNA is after peptide bond formation and before translocation. The P (peptidyl-tRNA) site - where peptidyl-tRNA is before peptide bond formation. The E (exit) site - where the uncharged tRNA from the P site goes after translocation. Peptidyl transferase - the active site that catalyzes formation of the peptide bond (23S rRNA in Prokaryotes) Polypeptide exit channel mRNA binding site The ribosome is a ribozyme?! “ …. most biologists did not seriously consider the possibility that RNA could be playing more than a bit part ... In the ribosome it has turned out that most of the intersubunit interface is RNA, the peptidyl transferase centre is RNA, and the decoding site and most of the A and P sites are RNA. It appears that the modern ribosome is composed of a somewhat geriatric, but functionally vital, RNA scaffold that is propped up and doted upon by its protein grandchildren. The ribosome Is one colossal enzyme.” James R. Williamson Scripps, Skaggs Inst. Nature 407: 306 Science 289:878 (T.R. Cech)Science289.878.pdf tRNAs Secondary Tertiary structure Isoacceptor tRNAs- Different tRNAs carrying the same aa tRNA identity- “2nd genetic code”- Special sequence elements on tRNAs that can be recognized by aaRS and then determine which aa is charged; Positive elements Negative elements Some special tRNAs Initiator tRNA: tRNAf Met tRNAiMet tmRNA Aminoacyl-tRNA Synthetases Two-step Reaction equation: 1) ATP + amino acid (AA) -- AMP-AA + PPi 2) tRNA + AMP-AA -- tRNA-AA + AMP Classification 1) Class I aaRSs 2) Class II aaRSs Proof-reading - quality c
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