GDH3 encodes a glutamate dehydrogenase isozyme, a previously unrecognized route for glutamate biosynthesis in Saccharomyces cerevisiae

被引:75
作者
Avendano, A [1 ]
Deluna, A [1 ]
Olivera, H [1 ]
Valenzuela, L [1 ]
Gonzalez, A [1 ]
机构
[1] NATL AUTONOMOUS UNIV MEXICO, INST FISIOL CELULAR, DEPT MICROBIOL, MEXICO CITY 04510, DF, MEXICO
关键词
D O I
10.1128/jb.179.17.5594-5597.1997
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
It has been considered that the yeast Saccharomyces cerevisiae, like many other microorganisms, synthesizes glutamate through the action of NADP(+)-glutamate dehydrogenase (NADP(+)-GDH), encoded by GDH1, or through the combined action of glutamine synthetase and glutamate synthase (GOGAT), encoded by GLN1 and GLT1, respectively. A double mutant of S cerevisiae lacking NADP(+)-GDN and GOGAT activities was constructed. This strain was able to grow on ammonium as the sole nitrogen source and thus to synthesize glutamate through an alternative pathway. A computer search for similarities between the GDH1 nucleotide sequence and the complete yeast genome was carried out. In addition to identifying its cognate sequence at chromosome XIV, the search found that GDH1 showed high identity with a previously recognized open reading frame (GDH3) of chromosome I. Triple mutants impaired in GDH1, GLT1, and GDH3 were obtained. These were strict glutamate auxotrophs. Our results indicate that GDH3 plays a significant physiological role, providing glutamate when GDH1 and GLT1 are impaired. This is the first example of a microorganism possessing three pathways for glutamate biosynthesis.
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页码:5594 / 5597
页数:4
相关论文
共 27 条
[1]  
[Anonymous], METHODS ENZYMOLOGY
[2]   ENZYME DEFECTS IN GLUTAMATE-REQUIRING STRAINS OF SCHIZOSACCHAROMYCES-POMBE [J].
BAREL, I ;
MACDONALD, DW .
FEMS MICROBIOLOGY LETTERS, 1993, 113 (03) :267-272
[3]   AMMONIUM ASSIMILATION IN RHIZOBIUM-PHASEOLI BY THE GLUTAMINE SYNTHETASE-GLUTAMATE SYNTHASE PATHWAY [J].
BRAVO, A ;
MORA, J .
JOURNAL OF BACTERIOLOGY, 1988, 170 (02) :980-984
[4]   THE NUCLEOTIDE-SEQUENCE OF CHROMOSOME-I FROM SACCHAROMYCES-CEREVISIAE [J].
BUSSEY, H ;
KABACK, DB ;
ZHONG, WW ;
VO, DT ;
CLARK, MW ;
FORTIN, N ;
HALL, J ;
OUELLETTE, BFF ;
KENG, T ;
BARTON, AB ;
SU, YP ;
DAVIES, CJ ;
STORMS, RK .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (09) :3809-3813
[5]   SACCHAROMYCES-CEREVISIAE HAS A SINGLE GLUTAMATE SYNTHASE GENE CODING FOR A PLANT-LIKE HIGH-MOLECULAR-WEIGHT POLYPEPTIDE [J].
COGONI, C ;
VALENZUELA, L ;
GONZALEZHALPHEN, D ;
OLIVERA, H ;
MACINO, G ;
BALLARIO, P ;
GONZALEZ, A .
JOURNAL OF BACTERIOLOGY, 1995, 177 (03) :792-798
[6]   UREIDOSUCCINIC ACID UPTAKE IN YEAST AND SOME ASPECTS OF ITS REGULATION [J].
DRILLIEN, R ;
LACROUTE, F .
JOURNAL OF BACTERIOLOGY, 1972, 109 (01) :203-&
[7]   ISOLATION AND CHARACTERIZATION OF A SACCHAROMYCES-CEREVISIAE MUTANT WITH IMPAIRED GLUTAMATE SYNTHASE ACTIVITY [J].
FOLCH, JL ;
ANTARAMIAN, A ;
RODRIGUEZ, L ;
BRAVO, A ;
BRUNNER, A ;
GONZALEZ, A .
JOURNAL OF BACTERIOLOGY, 1989, 171 (12) :6776-6781
[8]   WHY DOES ESCHERICHIA-COLI HAVE 2 PRIMARY PATHWAYS FOR SYNTHESIS OF GLUTAMATE [J].
HELLING, RB .
JOURNAL OF BACTERIOLOGY, 1994, 176 (15) :4664-4668
[9]   YEAST ESCHERICHIA-COLI SHUTTLE VECTORS WITH MULTIPLE UNIQUE RESTRICTION SITES [J].
HILL, JE ;
MYERS, AM ;
KOERNER, TJ ;
TZAGOLOFF, A .
YEAST, 1986, 2 (03) :163-167
[10]   TRANSFORMATION OF YEAST [J].
HINNEN, A ;
HICKS, JB ;
FINK, GR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1978, 75 (04) :1929-1933