D-ARABITOL METABOLISM IN CANDIDA-ALBICANS - CONSTRUCTION AND ANALYSIS OF MUTANTS LACKING D-ARABITOL DEHYDROGENASE

被引:26
作者
WONG, B
LEESON, S
GRINDLE, S
MAGEE, B
BROOKS, E
MAGEE, PT
机构
[1] UNIV CINCINNATI,DEPT CHEM,CINCINNATI,OH 45267
[2] UNIV MINNESOTA,DEPT GENET & CELL BIOL,ST PAUL,MN
关键词
D O I
10.1128/jb.177.11.2971-2976.1995
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Candida albicans produces large amounts of the acyclic pentitol D-arabitol in culture and in infected animals and humans, and most strains also grow on minimal D-arabitol medium, An earlier study showed that the major metabolic precursor of D-arabitol in C. albicans was D-ribulose-5-PO4, from the pentose pathway, that C. albicans contained ah NAD-dependent D-arabitol dehydrogenase (ArDH); and that the ArDH structural gene (ARD) encoded a 31-kDa short-chain dehydrogenase that catalyzed the reaction D-arabitol + NAD <=> D-ribulose + NADH. In the present study, we disrupted both ARD chromosomal alleles in C. albicans and analyzed the resulting mutants. The ard null mutation was verified by Southern hybridization, and the null mutant's inability to produce ArDH was verified by Western immunoblotting. The ard null mutant grew well on minimal glucose medium, brit it was unable to grow on minimal D-arabitol or D-arabinose medium. Thus, ArDH catalyzes the first step in D-arabitol utilization and a necessary intermediate step in D-arabinose utilization. Unexpectedly, the ard null mutant synthesized D-arabitol from glucose. Moreover, C-13 nuclear magnetic resonance studies showed that the ard null mutant and its wild-type parent synthesized D-arabitol via the same pathway. These results imply that C. albicans synthesizes and utilizes D-arabitol via separate metabolic pathways, which was not previously suspected for fungi.
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页码:2971 / 2976
页数:6
相关论文
共 29 条
[1]   A METHOD FOR GENE DISRUPTION THAT ALLOWS REPEATED USE OF URA3 SELECTION IN THE CONSTRUCTION OF MULTIPLY DISRUPTED YEAST STRAINS [J].
ALANI, E ;
CAO, L ;
KLECKNER, N .
GENETICS, 1987, 116 (04) :541-545
[2]   GPD1, WHICH ENCODES GLYCEROL-3-PHOSPHATE DEHYDROGENASE, IS ESSENTIAL FOR GROWTH UNDER OSMOTIC-STRESS IN SACCHAROMYCES-CEREVISIAE, AND ITS EXPRESSION IS REGULATED BY THE HIGH-OSMOLARITY GLYCEROL RESPONSE PATHWAY [J].
ALBERTYN, J ;
HOHMANN, S ;
THEVELEIN, JM ;
PRIOR, BA .
MOLECULAR AND CELLULAR BIOLOGY, 1994, 14 (06) :4135-4144
[3]  
BERNARD EM, 1982, J CLIN MICROBIOL, V146, P353
[4]   STUDIES ON FORMATION OF D-ARABITOL BY OSMOPHILIC YEASTS [J].
BLAKLEY, ER ;
SPENCER, JFT .
CANADIAN JOURNAL OF BIOCHEMISTRY AND PHYSIOLOGY, 1962, 40 (12) :1737-&
[5]   EXAMINATION OF PRODUCTION OF HYDROLYTIC ENZYMES AND TOXINS BY PATHOGENIC STRAINS OF CANDIDA-ALBICANS [J].
CHATTAWAY, FW ;
ODDS, FC ;
BARLOW, AJE .
JOURNAL OF GENERAL MICROBIOLOGY, 1971, 67 (AUG) :255-+
[6]  
FONZI WA, 1993, GENETICS, V134, P717
[7]   SERUM ARABINITOL CONCENTRATIONS AND ARABINITOL CREATININE RATIOS IN INVASIVE CANDIDIASIS [J].
GOLD, JWM ;
WONG, B ;
BERNARD, EM ;
KIEHN, TE ;
ARMSTRONG, D .
JOURNAL OF INFECTIOUS DISEASES, 1983, 147 (03) :504-513
[8]  
GORMAN JA, 1991, GENETICS, V129, P19
[9]   GENE ISOLATION BY COMPLEMENTATION IN CANDIDA-ALBICANS AND APPLICATIONS TO PHYSICAL AND GENETIC-MAPPING [J].
GOSHORN, AK ;
GRINDLE, SM ;
SCHERER, S .
INFECTION AND IMMUNITY, 1992, 60 (03) :876-884
[10]  
Guthrie C, 1991, GUIDE YEAST GENETICS