Structure of the yeast isoleucyl-tRNA synthetase gene (ILS1). DNA-sequence, amino-acid sequence of proteolytic peptides of the enzyme and comparison of the structure to those of other known aminoacyl-tRNA synthetases.
The ILS1 gene encoding for cytoplasmic isoleucyl-tRNA synthetase from Saccharomyces cerevisiae was subcloned from a 5.4-kb insert of the shuttle vector YEp13 to M13mp8 and M13mp9. Nucleotide sequence analysis of a 4.3-kb BamHI-HpaI fragment revealed a single open reading frame from which we deduced the amino-acid sequence of the enzyme. Independently obtained amino-acid sequence information from ten tryptic peptides of the purified enzyme confirmed the gene-derived structure. The enzyme is comprised of 1073 amino-acids consistent with earlier determinations of its molecular mass. The codon usage of ILS1 is typical of abundant yeast proteins. A significant homology to E. coli isoleucyl- and valyl-tRNA synthetases as well as to yeast valyl-tRNA synthetase was detected. The characteristic amino-acid residues of the aminoacyl-adenylate site and of the potential binding site of the 3'-end of tRNA found in other synthetases are present in the structure.
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Abteilung Chemie des Max-Planck-Institutes für experimentelle Medizin, Göttingen.
Englisch
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Markmeyer
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Schischkoff
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Sternbach
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Kratzin
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Cramer
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D000595:Amino Acid Sequence; D000604:Amino Acyl-tRNA Synthetases / Q000032:analysis* / Q000235:genetics; D001483:Base Sequence; D003201:Computers; D004271:DNA, Fungal / Q000032:analysis*; D004926:Escherichia coli / Q000235:genetics; D007533:Isoleucine-tRNA Ligase / Q000032:analysis* / Q000235:genetics; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics
D004271:DNA, Fungal; D000604:Amino Acyl-tRNA Synthetases; D007533:Isoleucine-tRNA Ligase