PMID:2405383
| Citation |
Leinfelder, W, Forchhammer, K, Veprek, B, Zehelein, E and Böck, A (1990) In vitro synthesis of selenocysteinyl-tRNA(UCA) from seryl-tRNA(UCA): involvement and characterization of the selD gene product. Proc. Natl. Acad. Sci. U.S.A. 87:543-7 |
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| Abstract |
The selD gene from Escherichia coli, whose product is involved in selenium metabolism, has been cloned and sequenced. selD codes for a protein of 347 amino acids with a calculated molecular weight of 36,687. Analysis of the selD gene product through expression of the gene in the phage T7 promoter/polymerase system confirmed the predicted molecular weight of the protein. Gene disruption experiments demonstrated that the SelD protein is required both for the incorporation of selenium into the modified nucleoside 5-methylaminomethyl-2-selenouridine of tRNA and for the biosynthesis of selenocysteine from an L-serine residue esterbonded to tRNA(Ser)(UCA). tRNA(Ser)(UCA) has been purified, aminoacylated with L-serine, and used as a substrate for the development of an in vitro system for selenocysteine biosynthesis. Efficient formation of selenocysteinyl-tRNA(Ser)(UCA) was achieved by using extracts in which both the selD and the selA gene products were overproduced. The results demonstrate that selenocysteine is synthesized from L-serine bound to tRNA(UCA) and they are in accord with SelD functioning as a donor of reduced selenium. |
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| Keywords |
Amino Acid Sequence; Bacterial Proteins/genetics; Base Sequence; Codon/genetics; Drosophila Proteins; Escherichia coli/genetics; Genes, Bacterial; Genotype; Molecular Sequence Data; Mutation; Phosphotransferases; Plasmids; RNA, Transfer, Amino Acid-Specific/metabolism; RNA, Transfer, Amino Acyl/biosynthesis; RNA, Transfer, Amino Acyl/genetics; RNA, Transfer, Ser/metabolism; Restriction Mapping; Selenium/metabolism |
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