title: SmpB triggers GTP hydrolysis of elongation factor Tu on ribosomes by compensating for the lack of codon-anticodon interaction during trans-translation initiation.
abstract: Bacterial tmRNA rescues ribosomes that stall because of defective mRNAs via the trans-translation process. Although entry of the charged transfer messenger RNA (tmRNA) into the ribosome proceeded in the absence of elongation factor (EF-Tu) and in the presence of EF-Tu and the antibiotic kirromycin, evidence was found for the involvement of EF-Tu in trans-translation initiation. The polyalanine synthesis system attained by using a tmRNA variant consisting of only the tRNA-like domain revealed that it was completely dependent on the presence of SmpB and greatly enhanced by EF-Tu and EF-G. Actually, ribosome-dependent GTPase activity of EF-Tu was stimulated by the addition of SmpB and tmRNA but independently of template mRNA, demonstrating that SmpB compensates for the lack of codon-anticodon interaction during the first step of the trans-translation initiation. Based on these results, we suggest that SmpB structurally mimics the anticodon arm of tRNA and elicits GTP hydrolysis of EF-Tu upon tmRNA accommodation in the A site of the ribosome.
authors.affiliation: Department of Medical Genome Sciences, Graduate School of Frontier Sciences, the University of Tokyo, FSB401, 5-1-5, Kashiwanoha, Kashiwa-shi, Chiba Prefecture 277-8562, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Shimizu
authors.forename: Takuya
authors.initials: T
authors.lastname: Ueda
mesh_terms: D000926:Anticodon; D003062:Codon; D006160:Guanosine Triphosphate / Q000378:metabolism*; D006868:Hydrolysis; D010444:Peptide Elongation Factor Tu / Q000378:metabolism*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D016601:RNA-Binding Proteins / Q000378:metabolism*
chemical_list: D000926:Anticodon; D003062:Codon; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C430328:small protein B; D006160:Guanosine Triphosphate; D010444:Peptide Elongation Factor Tu