Characterization of the cydAB-encoded cytochrome bd oxidase from Mycobacterium smegmatis

被引:125
作者
Kana, BD
Weinstein, EA
Avarbock, D
Dawes, SS
Rubin, H
Mizrahi, V
机构
[1] Univ Penn, Div Infect Dis, Philadelphia, PA 19104 USA
[2] S African Inst Med Res, MRC, SAIMR,WITS, Mol Mycobacteriol Res Unit, Johannesburg, South Africa
[3] Univ Witwatersrand, Sch Pathol, Dept Mol Med & Hematol, Johannesburg, South Africa
关键词
D O I
10.1128/JB.183.24.7076-7086.2001
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The cydAB genes from Mycobacterium smegmatis have been cloned and characterized. The cyd4 and cydB genes encode the two subunits of a cytochrome bd oxidase belonging to the widely distributed family of quinol oxidases found in prokaryotes. The cydD and cydC genes located immediately downstream of cydB encode a putative ATP-binding cassette-type transporter. At room temperature, reduced minus oxidized difference spectra of membranes purified from wild-type M. smegmatis displayed spectral features that are characteristic of the gamma -proteobacterial type cytochrome bd oxidase. Inactivation of cyd-4 or cydB by insertion of a kanamycin resistance marker resulted in loss of d-heme absorbance at 631 run. The d-heme could be restored by transformation of the M. smegmatis cyd mutants with a replicating plasmid carrying the highly homologous cydABDC gene cluster from Mycobacterium tuberculosis. Inactivation of cyd4 had no effect on the ability of M. smegmatis to exit from stationary phase at 37 or 42 degreesC. The growth. rate of the cydA mutant was tested under oxystatic conditions. Although no discernible growth defect was observed under moderately aerobic conditions (9.2 to 37.5 x 10(2) Pa of pO(2) or 5 to 21% air saturation), the mutant displayed a significant growth disadvantage hen cocultured with the wild type under extreme microaerophilia (0.8 to 1.7 x 10(2) Pa of pO(2) or 0.5 to 1% air saturation). These observations were in accordance with the two- to threefold increase in cydAB gene expression observed upon reduction of the pO(2), of the growth medium from 21 to 0.5% air saturation and with the concomitant increase in d-heme absorbance in spectra of membranes isolated from wild-type M. smegmatis cultured at 1% air saturation. Finally, the cydA mutant displayed a competitive growth disadvantage in the presence of the terminal oxidase inhibitor, cyanide, when cocultured with wild type at 21% air saturation in an oxystat. In conjunction with these findings, our results suggest that cytochrome bd is an important terminal oxidase in M. smegmatis.
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页码:7076 / 7086
页数:11
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共 66 条
[1]  
ASANO A, 1965, J BIOL CHEM, V240, P4002
[2]   A cytochrome bb′-type quinol oxidase in Bacillus subtilis strain 168 [J].
Azarkina, N ;
Siletsky, S ;
Borisov, V ;
von Wachenfeldt, C ;
Hederstedt, L ;
Konstantinov, AA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (46) :32810-32817
[3]   Allelic exchange in Mycobacterium tuberculosis with long linear recombination substrates [J].
Balasubramanian, V ;
Pavelka, MS ;
Bardarov, SS ;
Martin, J ;
Weisbrod, TR ;
McAdam, RA ;
Bloom, BR ;
Jacobs, WR .
JOURNAL OF BACTERIOLOGY, 1996, 178 (01) :273-279
[4]   Transmission of tuberculosis among the urban homeless [J].
Barnes, PF ;
ElHajj, H ;
PrestonMartin, S ;
Cave, MD ;
Jones, BE ;
Otaya, M ;
Pogoda, J ;
Eisenach, KD .
JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION, 1996, 275 (04) :305-307
[5]   Regulatory O-2 tensions for the synthesis of fermentation products in Escherichia coli and relation to aerobic respiration [J].
Becker, S ;
Vlad, D ;
Schuster, S ;
Pfeiffer, P ;
Unden, G .
ARCHIVES OF MICROBIOLOGY, 1997, 168 (04) :290-296
[6]   Purification, gene cloning, targeted knockout, overexpression, and biochemical characterization of the major pyrazinamidase from Mycobacterium smegmatis [J].
Boshoff, HIM ;
Mizrahi, V .
JOURNAL OF BACTERIOLOGY, 1998, 180 (22) :5809-5814
[7]   Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence [J].
Cole, ST ;
Brosch, R ;
Parkhill, J ;
Garnier, T ;
Churcher, C ;
Harris, D ;
Gordon, SV ;
Eiglmeier, K ;
Gas, S ;
Barry, CE ;
Tekaia, F ;
Badcock, K ;
Basham, D ;
Brown, D ;
Chillingworth, T ;
Connor, R ;
Davies, R ;
Devlin, K ;
Feltwell, T ;
Gentles, S ;
Hamlin, N ;
Holroyd, S ;
Hornby, T ;
Jagels, K ;
Krogh, A ;
McLean, J ;
Moule, S ;
Murphy, L ;
Oliver, K ;
Osborne, J ;
Quail, MA ;
Rajandream, MA ;
Rogers, J ;
Rutter, S ;
Seeger, K ;
Skelton, J ;
Squares, R ;
Squares, S ;
Sulston, JE ;
Taylor, K ;
Whitehead, S ;
Barrell, BG .
NATURE, 1998, 393 (6685) :537-+
[8]   Aerobic regulation of cytochrome d oxidase (cydAB) operon expression in Escherichia coli: Roles of Fnr and ArcA in repression and activation [J].
Cotter, PA ;
Melville, SB ;
Albrecht, JA ;
Gunsalus, RP .
MOLECULAR MICROBIOLOGY, 1997, 25 (03) :605-615
[9]   Mycobacterial stationary phase induced by low oxygen tension:: Cell wall thickening and localization of the 16-kilodalton α-crystallin homology [J].
Cunningham, AF ;
Spreadbury, CL .
JOURNAL OF BACTERIOLOGY, 1998, 180 (04) :801-808
[10]   The cioAB genes from Pseudomonas aeruginosa code for a novel cyanide-insensitive terminal oxidase related to the cytochrome bd quinol oxidases [J].
Cunningham, L ;
Pitt, M ;
Williams, HD .
MOLECULAR MICROBIOLOGY, 1997, 24 (03) :579-591