title: Antideterminants present in minihelix(Sec) hinder its recognition by prokaryotic elongation factor Tu.
abstract: During protein biosynthesis, all aminoacylated elongator tRNAs except selenocysteine-inserting tRNA Sec form ternary complexes with activated elongation factor. tRNA Sec is bound by its own translation factor, an elongation factor analogue, e.g. the SELB factor in prokaryotes. An apparent reason for this discrimination could be related to the unusual length of tRNA Sec amino acid-acceptor branch formed by 13 bp. However, it has been recently shown that an aspartylated minihelix of 13 bp derived from yeast tRNA Asp is an efficient substrate for Thermus thermophilus EF-Tu-GTP, suggesting that features other than the length of tRNA Sec prevent its recognition by EF-Tu-GTP. A stepwise mutational analysis of a minihelix derived from tRNA Sec in which sequence elements of tRNA Asp were introduced showed that the sequence of the amino acid- acceptor branch of Escherichia coli tRNA Sec contains a specific structural element that hinders its binding to T.thermophilus EF-Tu-GTP. This antideterminant is located in the 8th, 9th and 10th bp in the acceptor branch of tRNA Sec, corresponding to the last base pair in the amino acid acceptor stem and the two first pairs in the T-stem. The function of this C7.G66/G49.U65/C50.G64 box was tested by its transplantation into a minihelix derived from tRNA Asp, abolishing its recognition by EF-Tu-GTP. The specific role of this nucleotide combination is further supported by its absence in all known prokaryotic elongator tRNAs.
authors.affiliation: Institut de Biologie Moléculaire et Cellulaire du Centre National de la Recherche Scientifique, Strasbourg, France.
authors.forename: J
authors.initials: J
authors.lastname: Rudinger
authors.forename: R
authors.initials: R
authors.lastname: Hillenbrandt
authors.forename: M
authors.initials: M
authors.lastname: Sprinzl
authors.forename: R
authors.initials: R
authors.lastname: Giegé
mesh_terms: D001483:Base Sequence; D005075:Biological Evolution; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010444:Peptide Elongation Factor Tu / Q000378:metabolism*; D016372:RNA, Antisense / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012345:RNA, Transfer, Amino Acid-Specific / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012349:RNA, Transfer, Asp / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D016963:Thermus thermophilus / Q000378:metabolism
chemical_list: D016372:RNA, Antisense; D012345:RNA, Transfer, Amino Acid-Specific; D012349:RNA, Transfer, Asp; C036928:tRNA, selenocysteine-; D006160:Guanosine Triphosphate; D010444:Peptide Elongation Factor Tu