In vitro incorporation of selenium into tRNAs of Salmonella typhimurium

Zsuzsa Veres, Lin Tsai, Michael Politino, Thressa C. Stadtman

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Broken-cell preparations of Salmonella typhimurium rapidly incorporated 75Se from 75SeO32- into tRNA by an ATP-dependent process. Selenium incorporation in the presence of 50 μM 75SeO32- (0.8-1 pmol per A260 unit) was enhanced by the selenocysteine precursor, O-acetyl-L-serine (to 3.7 pmol per A260 unit). This increase in incorporation was a function of O-acetyl-L-serine concentration. Neither O-acetyl-L-homoserine nor O-phospho-L-serine stimulated the incorporation of selenium into tRNA. The incorporation of 75Se from 75SeO32- was decreased by adding L-selenocysteine but not by adding the D isomer. When homologous bulk tRNA was added to the broken-cell preparations, an increased rate of 75Se labeling was observed. The supernatant fraction of the broken-cell preparation contained all of the enzymes required for this process. Reversed-phase HPLC analysis of labeled bulk tRNA digested to nucleosides showed the presence of a labeled compound that coeluted with authentic 5-methylaminomethyl-2-selenouridine.

Original languageEnglish
Pages (from-to)6341-6344
Number of pages4
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number16
DOIs
Publication statusPublished - Aug 1990

Keywords

  • 5-methylaminomethyl-2-selenouridine
  • L-selenocysteine
  • Selenonucleoside

ASJC Scopus subject areas

  • General

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