High guanine plus cytosine content in the third letter of codons of an extreme thermophile. DNA sequence of the isopropylmalate dehydrogenase of Thermus thermophilus.
In studies on the cause of the extreme stability of the macromolecules of Thermus thermophilus HB8, the leuB gene coding for 3-isopropylmalate dehydrogenase of the leucine synthesis pathway and its flanking regions were cloned and sequenced. The leuB gene of T. thermophilus was expressed in a leuB-less mutant of Escherichia coli, and thermostable dehydrogenase was purified from an extract of the cells. The primary structure of the thermophilic isopropylmalate dehydrogenase was deduced from the nucleotide sequence leuB gene (1017 base pairs) and the amino acid sequence of the peptides isolated from the purified dehydrogenase. The thermophilic dehydrogenase has Mr = 35,968, and the value was close to that determined for the monomer of dehydrogenase (36,000) by gel electrophoresis. The molecular weight of active dimeric dehydrogenase was found to be 73,000 by high speed liquid chromatography. The primary structure of dehydrogenase was consistent with the amino acid composition of the dehydrogenase. In contrast to the isopropylmalate dehydrogenase of E. coli which contains 8 cysteine residues, there was no cysteine in thermophilic isopropylmalate dehydrogenase. The 5'-noncoding region contained a typical Shine-Dalgarno sequence. The guanine plus cytosine content of the coding region was 70.1%, and that of the third letter of the codons was extremely high (89.4%).
D050539:3-Isopropylmalate Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000235:genetics*; D000595:Amino Acid Sequence; D001482:Base Composition; D001483:Base Sequence; D003062:Codon / Q000235:genetics*; D003596:Cytosine / Q000032:analysis*; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D006147:Guanine / Q000032:analysis*; D012333:RNA, Messenger / Q000235:genetics*; D013824:Thermus / Q000201:enzymology / Q000235:genetics*
D003062:Codon; D004269:DNA, Bacterial; D012333:RNA, Messenger; D006147:Guanine; D003596:Cytosine; D000429:Alcohol Oxidoreductases; D050539:3-Isopropylmalate Dehydrogenase; D004262:DNA Restriction Enzymes