Localization of the binding site for the 3'-terminal sequence of tRNAPhe in subunits of phenylalanyl-tRNA synthetase from Thermus thermophilus

• Published in: Biochemistry (Moscow) Vol. 63; no. 9; pp. 1051 - 1056
• Main Authors: Moor, N A, Ankilova, V N, Favre, A, Lavrik, O I
• Format: Journal Article
• Language: English
• Published: United States 01.09.1998
• Subjects: Base Sequence; Binding Sites - genetics; Escherichia coli - genetics; Molecular Sequence Data; Nucleic Acid Conformation; Phenylalanine-tRNA Ligase - chemistry; Phenylalanine-tRNA Ligase - metabolism; Photoaffinity Labels; Protein Conformation; RNA, Bacterial - chemistry; RNA, Bacterial - genetics; RNA, Bacterial - metabolism; RNA, Transfer, Phe - chemistry; RNA, Transfer, Phe - genetics; RNA, Transfer, Phe - metabolism; Thermus thermophilus - enzymology; Thermus thermophilus - genetics; Thermus thermophilus - metabolism; Thiouridine - chemistry
• ISSN: 0006-2979

A photoreactive tRNAPhe derivative containing a 4-thiouridine residue at the 3'-end (tRNAPhe-s4U-75) was prepared by tRNA nucleotidyltransferase-mediated incorporation of s4UMP into a tRNAPhe transcript lacking the 3'-terminal dinucleotide. The resulting tRNAPhe-s4U-75 was covalently bound to phenylalanyl-tRNA synthetase from Thermus thermophilus, and all criteria of an affinity modification were met. The main products of modification displaying various electrophoretic mobilities were formed by binding tRNAPhe-s4U-75 to the beta-subunit (major) of the enzyme. These data suggest that the nucleotide found at position 75 of tRNAPhe interacts with the beta-subunit of phenylalanyl-tRNA synthetase.