Sequence and expression of the rusticyanin structural gene from Thiobacillus ferrooxidans ATCC33020 strain.
The periplasmic blue copper protein rusticyanin is thought to play an important role in iron oxidation by Thiobacillus ferrooxidans. We present the sequence of the gene, rus, encoding rusticyanin together with about 1.4 kb of upstream and 0.3 kb of downstream DNA. The rus gene is unique to T. ferrooxidans. Evidence is presented that it is the last gene of an operon and that it can be transcribed from its own promoter. In ATCC33020 strain, rusticyanin is synthesized in ferrous iron but also in sulfur growth conditions suggesting that it could play a role in both energetic metabolisms. The rus gene transcribed from a vector promoter in Escherichia coli leads to the production of a processed aporusticyanin in the periplasmic space, indicating that its signal sequence is correctly recognized by the secretion machinery and the signal peptidase of E. coli.
Bengrine
A
A
Laboratoire de Chimie Bactérienne, Institut de Biologie Structurale et de Microbiologie, Centre National de la Recherche Scientifique, 31 chemin Joseph Aiguier, 13402 Marseille Cedex 20, France.
Guiliani
N
N
Appia-Ayme
C
C
Jedlicki
E
E
Holmes
D S
DS
Chippaux
M
M
Bonnefoy
V
V
D000595:Amino Acid Sequence; D001400:Azurin / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction / Q000379:methods; D017421:Sequence Analysis; D013855:Thiobacillus / Q000235:genetics*; D014158:Transcription, Genetic
D001426:Bacterial Proteins; C012543:rusticyanin; D001400:Azurin