Mercuric reductase structural genes from plasmid R100 and transposon Tn501: functional domains of the enzyme.
The nucleotide sequence for the 2240 bp of plasmid R100 following the merC gene of the mercuric resistance operon has been determined and compared with the homologous sequence of transposon Tn501. The sequences following merC and preceding the next structural gene merA are unrelated between R100 and Tn501 and differ in length, with 72 bp in Tn501 and 509 bp in R100. The R100 sequence has a potential open reading frame (ORF) for a 140 amino acid polypeptide with a reasonable translational start signal preceding it. The merA genes contain 1686 (Tn501) and 1695 (R100) bp respectively. When optimally aligned, the merA sequences differ in 18% of their positions. These differences were clustered in specific regions. In addition, there was one nucleotide triplet in the Tn501 sequence which has no counterpart in the R100 sequence and one dodecyl-nucleotide sequence in the R100 sequence without counterpart in Tn501. Thus the predicted merA polypeptide of Tn501 contains 561 amino acids and the R100 counterpart contains 564 amino acids. Comparison of the R100 mercuric reductase sequences with that for human glutathione reductase [Krauth-Siegel et al.: Eur. J. Biochem. 121 (1982) 259-267], for which there is a 2 A resolution electron density map [Thieme et al.: J. Mol. Biol. 152 (1981) 763-782] shows a strong homology, with 26% identical amino acids and many conservative substitutions. This homology allows the conclusion that the active site of these enzymes and the contact positions for flavin adenine dinucleotide (FAD) and NADPH are highly conserved, while the amino- and carboxyl-terminal sequences differ.
Misra
T K
TK
Brown
N L
NL
Haberstroh
L
L
Schmidt
A
A
Goddette
D
D
Silver
S
S
D000595:Amino Acid Sequence; D001483:Base Sequence; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D008058:Dihydrolipoamide Dehydrogenase / Q000235:genetics; D005796:Genes; D005798:Genes, Bacterial; D005980:Glutathione Reductase / Q000235:genetics; D008628:Mercury / Q000633:toxicity*; D009876:Operon; D010088:Oxidoreductases / Q000235:genetics* / Q000276:immunology; D010957:Plasmids; D011550:Pseudomonas aeruginosa / Q000235:genetics; D012763:Shigella flexneri / Q000235:genetics
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D010088:Oxidoreductases; C021551:mercuric reductase; D008058:Dihydrolipoamide Dehydrogenase; D005980:Glutathione Reductase; D008628:Mercury