SACCHAROMYCES-CEREVISIAE HAS A SINGLE GLUTAMATE SYNTHASE GENE CODING FOR A PLANT-LIKE HIGH-MOLECULAR-WEIGHT POLYPEPTIDE

被引:41
作者
COGONI, C
VALENZUELA, L
GONZALEZHALPHEN, D
OLIVERA, H
MACINO, G
BALLARIO, P
GONZALEZ, A
机构
[1] Univ Nacl Autonoma Mexico, INST FISIOL CELULAR, DEPT MICROBIOL, MEXICO CITY 04510, DF, MEXICO
[2] Univ Nacl Autonoma Mexico, INST FISIOL CELULAR, DEPT BIOENERGET, MEXICO CITY 04510, DF, MEXICO
[3] UNIV ROMA LA SAPIENZA, POLICLIN UMBERTO 1, DIPARTIMENTO BIOPATOL UMANA, ROME, ITALY
[4] UNIV ROMA LA SAPIENZA, CTR STUDIO ACIDI NUCLEICI, DIPARTIMENTO GENET & BIOL MOLEC, ROME, ITALY
关键词
D O I
10.1128/jb.177.3.792-798.1995
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Purification of the glutamate synthase (GOGAT) enzyme from Saccharomyces cerevisiae showed that it is an oligomeric enzyme composed of three identical 199-kDa subunits. The GOGAT structural gene was isolated by screening a yeast genomic library with a yeast PCR probe. This probe was obtained by amplification with degenerate oligonucleotides designed from conserved regions of known GOGAT genes, The derived aminoterminal sequence of the GOGAT gene was confirmed by direct amino-terminal sequence analysis of the purified protein of 199 kDa. Northern (RNA) analysis allowed the identification of an mRNA of about 7 or 8 kb. An internal fragment of the GOGAT gene was used to obtain null GOGAT mutants completely devoid of GOGAT activity. The results show that S. cerevisiae has a single NADH-GOGAT enzyme, consisting of three 199-kDa monomers, that differs from the one found in prokaryotic microorganisms but is similar to those found in other eukaryotic organisms such as alfalfa.
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收藏
页码:792 / 798
页数:7
相关论文
共 44 条
[1]   PURIFICATION AND CHARACTERIZATION OF NADH-GLUTAMATE SYNTHASE FROM ALFALFA ROOT-NODULES [J].
ANDERSON, MP ;
VANCE, CP ;
HEICHEL, GH ;
MILLER, SS .
PLANT PHYSIOLOGY, 1989, 90 (01) :351-358
[2]  
AVILA C, 1993, PLANTA, V189, P475, DOI 10.1007/BF00198209
[3]  
BASTARRACHEA F, 1980, GLUTAMINE METABOLISM, P213
[4]   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
[5]  
BRUNO AO, 1979, METHOD ENZYMOL, V63, P159
[6]   2 ISOENZYMES OF NADH-DEPENDENT GLUTAMATE SYNTHASE IN ROOT-NODULES OF PHASEOLUS-VULGARIS L - PURIFICATION, PROPERTIES AND ACTIVITY CHANGES DURING NODULE DEVELOPMENT [J].
CHEN, FL ;
CULLIMORE, JV .
PLANT PHYSIOLOGY, 1988, 88 (04) :1411-1417
[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]   MOLECULAR-CLONING OF THE ACTIN GENE FROM YEAST SACCHAROMYCES CEREVISIAE [J].
GALLWITZ, D ;
SEIDEL, R .
NUCLEIC ACIDS RESEARCH, 1980, 8 (05) :1043-1059
[9]   STRUCTURAL ORGANIZATION OF THE GENES THAT ENCODE 2 GLUTAMATE SYNTHASE SUBUNITS OF ESCHERICHIA-COLI [J].
GARCIARRUBIO, A ;
LOZOYA, E ;
COVARRUBIAS, A ;
BOLIVAR, F .
GENE, 1983, 26 (2-3) :165-170
[10]   CLONING OF A YEAST GENE CODING FOR THE GLUTAMATE SYNTHASE SMALL SUBUNIT (GUS2) BY COMPLEMENTATION OF SACCHAROMYCES-CEREVISIAE AND ESCHERICHIA-COLI GLUTAMATE AUXOTROPHS [J].
GONZALEZ, A ;
MEMBRILLOHERNANDEZ, J ;
OLIVERA, H ;
ARANDA, C ;
MACINO, G ;
BALLARIO, P .
MOLECULAR MICROBIOLOGY, 1992, 6 (03) :301-308