Mirror image alternative interaction patterns of the same tRNA with either class I arginyl-tRNA synthetase or class II aspartyl-tRNA synthetase.
Gene cloning, overproduction and an efficient purification protocol of yeast arginyl-tRNA synthetase (ArgRS) as well as the interaction patterns of this protein with cognate tRNAArgand non-cognate tRNAAspare described. This work was motivated by the fact that the in vitro transcript of tRNAAspis of dual aminoacylation specificity and is not only aspartylated but also efficiently arginylated. The crystal structure of the complex between class II aspartyl-tRNA synthetase (AspRS) and tRNAAsp, as well as early biochemical data, have shown that tRNAAspis recognized by its variable region side. Here we show by footprinting with enzymatic and chemical probes that transcribed tRNAAspis contacted by class I ArgRS along the opposite D arm side, as is homologous tRNAArg, but with idiosyncratic interaction patterns. Besides protection, footprints also show enhanced accessibility of the tRNAs to the structural probes, indicative of conformational changes in the complexed tRNAs. These different patterns are interpreted in relation to the alternative arginine identity sets found in the anticodon loops of tRNAArgand tRNAAsp. The mirror image alternative interaction patterns of unmodified tRNAAspwith either class I ArgRS or class II AspRS, accounting for the dual identity of this tRNA, are discussed in relation to the class defining features of the synthetases. This study indicates that complex formation between unmodified tRNAAspand either ArgRS and AspRS is solely governed by the proteins.
Sissler
M
M
Unité Propre de Recherche 9002 'Structure des Macromolécules Biologiques et Mécanismes de Reconnaissance', Institut de Biologie Moléculaire et Cellulaire du Centre National de la Recherche Scientifique, Strasbourg, France.
Eriani
G
G
Martin
F
F
Giegé
R
R
Florentz
C
C
D000926:Anticodon / Q000737:chemistry; D001126:Arginine-tRNA Ligase / Q000145:classification / Q000378:metabolism*; D001226:Aspartate-tRNA Ligase / Q000145:classification / Q000378:metabolism*; D001483:Base Sequence; D018983:DNA Footprinting; D004926:Escherichia coli; D005656:Fungal Proteins / Q000145:classification / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D012331:RNA, Fungal / Q000737:chemistry / Q000378:metabolism*; D012347:RNA, Transfer, Arg / Q000737:chemistry / Q000378:metabolism*; D012349:RNA, Transfer, Asp / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity
D000926:Anticodon; D005656:Fungal Proteins; D012331:RNA, Fungal; D012347:RNA, Transfer, Arg; D012349:RNA, Transfer, Asp; D011993:Recombinant Fusion Proteins; D001226:Aspartate-tRNA Ligase; D001126:Arginine-tRNA Ligase