The contacts of yeast tRNA(Ser) with seryl-tRNA synthetase studied by footprinting experiments.
Yeast tRNA(Ser) is a member of the class II tRNAs, whose characteristic is the presence of an extended variable loop. This additional structural feature raises questions about the recognition of these class II tRNAs by their cognate synthetase and the possibility of the involvement of the extra arm in the recognition process. A footprinting study of yeast tRNA(Ser) complexed with its cognate synthetase, yeast seryl-tRNA synthetase (an alpha 2 dimer), was undertaken. Chemical (ethylnitrosourea) and enzymatic (nucleases S1 and V1) probes were used in the experiments. A map of the contact points between the tRNA and the synthetase was established and results were analyzed with respect to a three-dimensional model of yeast tRNA(Ser). Regions in close vicinity with the synthetase are clustered on one face of tRNA. The extra arm, which is strongly protected from chemical modifications, appears as an essential part of the contact area. The anticodon triplet and a large part of the anticodon arm are, in contrast, still accessible to the probes when the complex is formed. These results are discussed in the context of the recognition of tRNAs in the aminoacylation reaction.
Dock-Bregeon
A C
AC
Laboratoires de Biochimie et de Cristallographie, Centre National de la Recherche Scientifique, Strasbourg, France.
Garcia
A
A
Giegé
R
R
Moras
D
D
D000604:Amino Acyl-tRNA Synthetases / Q000037:antagonists & inhibitors*; D000926:Anticodon; D001345:Autoradiography; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D004722:Endoribonucleases; D005038:Ethylnitrosourea; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010449:Peptide Mapping; D012346:RNA, Transfer, Amino Acyl / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D012714:Serine-tRNA Ligase / Q000032:analysis / Q000037:antagonists & inhibitors*; D015719:Single-Strand Specific DNA and RNA Endonucleases
D000926:Anticodon; D012346:RNA, Transfer, Amino Acyl; D004722:Endoribonucleases; C032825:ribonuclease V(1); D015719:Single-Strand Specific DNA and RNA Endonucleases; D000604:Amino Acyl-tRNA Synthetases; D012714:Serine-tRNA Ligase; D005038:Ethylnitrosourea