New photoactivatable structural and affinity probes of RNAs: specific features and applications for mapping of spermine binding sites in yeast tRNA(Asp) and interaction of this tRNA with yeast aspartyl-tRNA synthetase.
Aryldiazonium salts are shown to be useful as phototriggered structural probes for RNA mapping as well as for footprinting of RNA/protein interaction. In particular the yeast tRNA(Asp)/aspartyl-tRNA synthetase complex is shown to involve the variable loop face and the concave side of the L-shaped nucleic acid bound to a lipophilic area of the enzyme. When chemically linked to spermine, the photoactive group cleaves RNA at polyamine binding sites; 3-4 spermines have been located in the tRNA(Asp), stabilizing the central part of the molecule in regions where two ribose-phosphate strands are close to each other.
Laboratoire de Biochimie, Institut de Biologie Moléculaire et Cellulaire du CNRS, Strasbourg, France.
A
A
Garcia
R
R
Giegé
J P
JP
Behr
D000345:Affinity Labels; D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism*; D001226:Aspartate-tRNA Ligase / Q000378:metabolism*; D001665:Binding Sites; D066298:In Vitro Techniques; D009690:Nucleic Acid Conformation; D010777:Photochemistry; D012345:RNA, Transfer, Amino Acid-Specific / Q000378:metabolism*; D012349:RNA, Transfer, Asp / Q000378:metabolism* / Q000648:ultrastructure; D012441:Saccharomyces cerevisiae; D013096:Spermine / Q000378:metabolism*
D000345:Affinity Labels; D012345:RNA, Transfer, Amino Acid-Specific; D012349:RNA, Transfer, Asp; D013096:Spermine; D000604:Amino Acyl-tRNA Synthetases; D001226:Aspartate-tRNA Ligase