Cloning and characterization of the azurin iso-1 gene, concerned with the electron transport chain involved in methylamine/methanol oxidation in the obligate methylotroph Methylomonas sp. strain J.
Two azurin-type blue copper proteins, which is concerned with the electron transport chain involved in methylamine/methanol oxidation, have been found in the obligate methylotroph Methylomonas sp. strain J. The azurin iso-1 gene was cloned and sequenced to analyze the role in the electron transport chain. PCR products synthesized with primers based on the N- and C-terminal amino acid sequences of azurin iso-1 were used as probes for cloning. One complete open reading frame (the azurin iso-1 gene) and one partial orf (orf1) were found in a cloned Eco105I-HindIII fragment, pMAZ3, with a total of 1066 bp. The gene encoded 148 amino acid residues. The amino acid sequence after Ala-21, deduced from the nucleotide sequence, was identical to that of the azurin iso-1 protein. The gene was in a region separate from the mau gene cluster in the chromosome. Escherichia coli expressed azurin iso-1. The results of northern blotting analysis suggested that expression of the azurin iso-1 gene is regulated by a complex regulatory network controlling oxidation of methylamine or methanol in this strain; for example, copper ions affected the expression of the azurin iso-1 gene.
Taguchi
K
K
Laboratory of Biochemistry, College of Science, Rikkyo (St. Paul's) University, Tokyo, Japan.
Kudo
T
T
Tobari
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001400:Azurin / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D015342:DNA Probes; D004579:Electron Transport; D015964:Gene Expression Regulation, Bacterial; D000432:Methanol / Q000378:metabolism*; D008744:Methylamines / Q000378:metabolism*; D008772:Methylococcaceae / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D016133:Polymerase Chain Reaction
D015342:DNA Probes; D008744:Methylamines; D001400:Azurin; D000432:Methanol