mRNA作用.pptVIP

Translation initiation - big picture In the absence of mRNA, ribosomes are dissociated Translation is initiated by recruitment of f-Met-tRNAMet and mRNA to the small ribosomal subunit Recruitment is facilitated by a number of initiation factors Initiation is complex in eukaryotes but simpler in prokaryotes There are many more levels of control in eukaryotes Initiation prokaryotes vs. eukaryotes Translation initiation in prokaryotes There are few Initiation factors IF1 (7.1 kDa) binds after ribosome dissociation, binds to A-site of small subunit assists IF2 and IF3 IF2 (79.7 kDa), homologous to eIF5B binds fMet-tRNAi GTPase IF3 (20.7 kDa) ribosome anti-association Involved in tRNAi-codon interaction, decoding Assures together with IF2 that fMet-tRNAini is bound not any other W2 (71 kDa), ATPase, eIF4A (helicase) homolog? Little known, no solid evidence for helicase function, stimulates in-vitro translation of some mRNA with secondary structure around AUG EF-P (21 kDa), eIF5A homolog Little known Model for Initiation in Prokaryotes Dissociation of ribosome due to absence of mRNA and aided by IF3 binding to 30S IF3 was the earliest- and most-studied bacterial initiation factor Association to 30S of (which order is unknown) IF1 into A-site ternary complex (IF2, fMet-tRNAi, GTP) - places fMet-tRNAi into P-site mRNA at AUG codon (Shine Dalgarno) - decoding task - get the AUG codon matching the anticodon of fMet-tRNAMet - GUG(8%) and UUG (1%) are sometimes used - internal AUGs vs initiation codons This is followed by dissociation of IF1 and IF3 50S particle joins IF2 is released with GTP hydrolysis (what is the GEF for IF2?) Elongation starts What is known structurally? IF1 (eIF1A) has OB-fold and has been located on the 30S particle IF2 (eIF5B) structure solved and approximate placement on 30S has been mapped with footprinting experiments IF3(no eukaryotic homolog) N- and C-terminal domains known and partial structures on 30S particle Role of IF3 and initial