Identification of a functional homolog of the Escherichia coli and Salmonella typhimurium cysM gene encoding O-acetylserine sulfhydrylase B in Campylobacter jejuni.
The final step of L-cysteine biosynthesis in Escherichia coli and Salmonella typhimurium consists of the formation of L-cysteine from O-acetylserine and sulfide. This reaction can be catalyzed by two enzymes, O-acetylserine sulfhydrylase A and O-acetylserine sulfhydrylase B, the former of which has been more rigorously characterized. In contrast to O-acetylserine sulfhydrylase A, O-acetylserine sulfhydrylase B is preferentially used for cysteine biosynthesis during anaerobic growth and is able to utilize thiosulfate as a substrate. Campylobacter jejuni is a micro-aerophilic, Gram-negative bacterium, and a member of the epsilon subdivision of eubacteria. We have cloned, sequenced, and expressed a gene from C. jejuni that encodes a protein of 299 aa with a calculated molecular mass of 32,367 Da. Complementation analysis of an E. coli cysteine auxotroph with the pMEK34-14 recombinant plasmid containing a 1.2-kb insert of chromosomal DNA from C. jejuni revealed that transformants were capable of growth in medium containing either sulfide or thiosulfate as sole sulfur sources. These data indicate that the cloned C. jejuni gene is a functional homolog of the cysM gene that codes for O-acetylserine sulfhydrylase B in E. coli and S. typhimurium.
Garvis
S G
SG
Washington State University, Department of Microbiology, Pullman 99164-4233, USA.
Tipton
S L
SL
Konkel
M E
ME
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D016123:Campylobacter jejuni / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D003545:Cysteine / Q000378:metabolism; D003547:Cysteine Synthase / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010957:Plasmids; D011995:Recombination, Genetic; D012486:Salmonella typhimurium / Q000235:genetics; D013440:Sulfides / Q000378:metabolism; D013885:Thiosulfates / Q000378:metabolism
D004269:DNA, Bacterial; D013440:Sulfides; D013885:Thiosulfates; D003547:Cysteine Synthase; D003545:Cysteine