Saccharomyces cerevisiae has a single glutamate synthase gene coding for a plant-like high-molecular-weight polypeptide
Por:
Cogoni C., Valenzuela L., Gonzalez-Halphen D., Olivera H., Macino G., Ballario P., Gonzalez A.
Publicada:
1 ene 1995
Resumen:
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 amino-terminal 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.
Filiaciones:
Cogoni C.:
Departamento de Microbiologia, UNAM, Instituto de Fisiologia Celular, Apartado Postal 70-242, Mexico City 04510, Mexico
Valenzuela L.:
Departamento de Microbiologia, UNAM, Instituto de Fisiologia Celular, Apartado Postal 70-242, Mexico City 04510, Mexico
Gonzalez-Halphen D.:
Departamento de Microbiologia, UNAM, Instituto de Fisiologia Celular, Apartado Postal 70-242, Mexico City 04510, Mexico
Olivera H.:
Departamento de Microbiologia, UNAM, Instituto de Fisiologia Celular, Apartado Postal 70-242, Mexico City 04510, Mexico
Macino G.:
Departamento de Microbiologia, UNAM, Instituto de Fisiologia Celular, Apartado Postal 70-242, Mexico City 04510, Mexico
Ballario P.:
Departamento de Microbiologia, UNAM, Instituto de Fisiologia Celular, Apartado Postal 70-242, Mexico City 04510, Mexico
Gonzalez A.:
Departamento de Microbiologia, UNAM, Instituto de Fisiologia Celular, Apartado Postal 70-242, Mexico City 04510, Mexico
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