Interaction of human phenylalanyl-tRNA synthetase with specific tRNA according to thiophosphate footprinting.
The interaction of human cytoplasmic phenylalanyl-tRNA synthetase (an enzyme with yet unknown 3D-structure) with homologous tRNA(Phe) under functional conditions was studied by footprinting based on iodine cleavage of thiophosphate-substituted tRNA transcripts. Most tRNA(Phe) nucleotides recognized by the enzyme in the anticodon (G34), anticodon stem (G30-C40, A31-U39), and D-loop (G20) have effectively or moderately protected phosphates. Other important specificity elements (A35 and A36) were found to form weak nonspecific contacts. The D-stem, T-arm, and acceptor stem are also among continuous contacts of the tRNA(Phe) backbone with the enzyme, thus suggesting the presence of additional recognition elements in these regions. The data indicate that mechanisms of interaction between phenylalanyl-tRNA synthetases and specific tRNAs are different in prokaryotes and eukaryotes.
Vasil'eva
I A
IA
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
Semenova
E A
EA
Moor
N A
NA
D000595:Amino Acid Sequence; D049148:Aminoacylation; D001483:Base Sequence; D006801:Humans; D006868:Hydrolysis; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010652:Phenylalanine-tRNA Ligase / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry*; D011487:Protein Conformation; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry*
D010710:Phosphates; D012346:RNA, Transfer, Amino Acyl; D010652:Phenylalanine-tRNA Ligase; C035638:thiophosphoric acid