Preparation of oligonucleotides corresponding to the acceptor stem of yeast tRNAPhe and their interaction with yeast ATP(CTP):tRNA nucleotidyltransferase.
Seven oligonucleotides corresponding to the 3' and 5' sequences of the acceptor stem of yeast tRNAPhe have been prepared by chemical synthesis, chemical-enzymatic synthesis or by isolation from tRNA hydrolysates. The oligonucleotides have been examined as substrates for phosphodiester bond synthesis in the presence of ATP as catalysed by yeast ATP (CTP): tRNA nucleotidyltransferase. Oligonucleotides which correspond to the sequence of the 3'-strand of the tRNA acceptor stem and possess no secondary structure exhibit little or no activity with the enzyme. The ability of the enzyme to catalyse the synthesis of a phosphodiester linkage using ATP and an oligonucleotide corresponding to the 3'-strand of the acceptor stem is in general dramatically increased when an oligonucleotide corresponding to the sequence of the 5'-strand of tRNA acceptor stem is present. In cases where significant activity was observed kinetic parameters have been determined.
D000249:Adenosine Monophosphate / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D007202:Indicators and Reagents; D008099:Liver / Q000201:enzymology; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000138:chemical synthesis*; D009843:Oligoribonucleotides / Q000138:chemical synthesis*; D012316:RNA Nucleotidyltransferases / Q000378:metabolism*; D012346:RNA, Transfer, Amino Acyl / Q000138:chemical synthesis*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D007202:Indicators and Reagents; D009841:Oligonucleotides; D009843:Oligoribonucleotides; D012346:RNA, Transfer, Amino Acyl; D000249:Adenosine Monophosphate; D012316:RNA Nucleotidyltransferases; C023198:tRNA nucleotidyltransferase