Cloning and expression analysis of the duplicated genes for carbon monoxide dehydrogenase of Mycobacterium sp. strain JC1 DSM 3803.
Carbon monoxide dehydrogenase (CO-DH) is an enzyme catalysing the oxidation of CO to carbon dioxide in Mycobacterium sp. strain JC1 DSM 3803. Cloning of the genes encoding CO-DH from the bacterium and sequencing of overlapping clones revealed the presence of duplicated sets of genes for three subunits of the enzyme, cutB1C1A1 and cutB2C2A2, in operons, and a cluster of genes encoding proteins that may be involved in CO metabolism, including a possible transcriptional regulator. Phylogenetic analysis based on the amino acid sequences of large subunits of CO-DH suggested that the CO-DHs of Mycobacterium sp. JC1 and other mycobacteria are distinct from those of other types of bacteria. The growth phenotype of mutant strains lacking cutA genes and of a corresponding complemented strain showed that both of the duplicated sets of CO-DH genes were functional in this bacterium. Transcriptional fusions of the cutB genes with lacZ revealed that the cutBCA operons were expressed regardless of the presence of CO and were further inducible by CO. Primer extension analysis indicated two promoters, one expressed in the absence of CO and the other induced in the presence of CO. This is believed to be the first report to show the presence of multiple copies of CO-DH genes with identical sequences and in close proximity in carboxydobacteria, and to present the genetic evidence for the function of the genes in mycobacteria.
Song
Taeksun
T
Genome Research Center for Respiratory Pathogens, Yonsei University College of Medicine, Seoul 120-752, Republic of Korea.|Department of Biology, Yonsei University, Seoul 120-749, Republic of Korea.
Park
Sae Woong
SW
Department of Biology, Yonsei University, Seoul 120-749, Republic of Korea.
Park
Su-Jeong
SJ
Department of Microbiology, Pusan National University, Busan 609-735, Republic of Korea.
Kim
Ji Hyang
JH
Department of Biology, Yonsei University, Seoul 120-749, Republic of Korea.
Yu
Ji Young
JY
Genome Research Center for Respiratory Pathogens, Yonsei University College of Medicine, Seoul 120-752, Republic of Korea.
Oh
Jeong-Il
JI
Department of Microbiology, Pusan National University, Busan 609-735, Republic of Korea.
Kim
Young M
YM
Department of Biology, Yonsei University, Seoul 120-749, Republic of Korea.
D000445:Aldehyde Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D020440:Gene Duplication; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics* / Q000378:metabolism; D009161:Mycobacterium / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D010802:Phylogeny
D001426:Bacterial Proteins; D009097:Multienzyme Complexes; D000445:Aldehyde Oxidoreductases; C019823:carbon monoxide dehydrogenase