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Abstract:

The role of glutamate as osmoprotector was investigated through the study of a mutation in its biosynthetic pathway. A glt::Tn917-lacZ-cat insertion mutant (N1) conferring glutamate auxotrophy and enhanced β-galactosidase expression on high-salt media was selected. Co-transformation experiments and PCR analysis allowed locating the insertion into the gltB gene corresponding to the small unit of the glutamate synthase (GOGAT). The N1 mutant strain presented a glutamate requirement for growth and a tenfold decrease in GOGAT activity. Transcriptional activity of GOGAT, measured as β-galactosidase from the transposon fusion, correlated with enzymatic activity; expression was enhanced at the stationary phase and in high-ionic-strength media. However, osmotolerance of cultures of N1 mutant were as wild-type (wt), at least in semi-rich medium. In contrast, sporulation was slightly reduced (75% of wt), and spores were less resistant to UV, heat, and osmolarity, properties linked to the content of small, acid-soluble proteins (SASP). The content of these proteins was, in fact, reduced, in particular the SASP-γ type. The peptidoglycan-cortex, however, was not impaired since spores maintained lysozyme resistance. Addition of glutamate during sporulation partially rescued spore resistance, but germination and outgrowth remained impaired. Deficiencies in germination and outgrowth were also observed with spores from a gltA mutant strain. Taken together, these results pointed to the importance of GOGAT activity during sporulation, in particular for the synthesis SASPs.

Registro:

Documento: Artículo
Título:Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties
Autor:Ruzal, S.M.; Sanchez-Rivas, C.
Filiación:Fac. de Ciencias Exactas y Naturales, Departamento de Quimica Biologica, Pabellon II, Ciudad Universitaria, (1428) Buenos Aires, Argentina
Palabras clave:bacterial protein; beta galactosidase; glutamate synthase; glutamic acid; lysozyme; article; Bacillus subtilis; bacterial mutation; bacterial spore; enzyme activity; germination; heat tolerance; nonhuman; priority journal; radiosensitivity; sporogenesis; transposon; Artificial Gene Fusion; Bacillus subtilis; Bacterial Proteins; beta-Galactosidase; DNA Transposable Elements; Gene Expression Regulation, Bacterial; Genes, Bacterial; Genes, Reporter; Glutamate Synthase; Glutamic Acid; Muramidase; Mutagenesis, Insertional; Osmotic Pressure; Peptidoglycan; Spores, Bacterial; Transcription, Genetic; Bacillus subtilis; Bacteria (microorganisms); Felis catus; insertion sequences; Posibacteria; transposons
Año:2003
Volumen:47
Número:3
Página de inicio:208
Página de fin:213
DOI: http://dx.doi.org/10.1007/s00284-002-3960-x
Título revista:Current Microbiology
Título revista abreviado:Curr. Microbiol.
ISSN:03438651
CODEN:CUMID
CAS:glutamic acid, 11070-68-1, 138-15-8, 56-86-0, 6899-05-4; lysozyme, 9001-63-2; Bacterial Proteins; beta-Galactosidase, EC 3.2.1.23; DNA Transposable Elements; Glutamate Synthase, EC 1.4.1.13; Glutamic Acid, 56-86-0; Muramidase, EC 3.2.1.17; Peptidoglycan; spore-specific proteins, Bacillus
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03438651_v47_n3_p208_Ruzal

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Citas:

---------- APA ----------
Ruzal, S.M. & Sanchez-Rivas, C. (2003) . Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties. Current Microbiology, 47(3), 208-213.
http://dx.doi.org/10.1007/s00284-002-3960-x
---------- CHICAGO ----------
Ruzal, S.M., Sanchez-Rivas, C. "Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties" . Current Microbiology 47, no. 3 (2003) : 208-213.
http://dx.doi.org/10.1007/s00284-002-3960-x
---------- MLA ----------
Ruzal, S.M., Sanchez-Rivas, C. "Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties" . Current Microbiology, vol. 47, no. 3, 2003, pp. 208-213.
http://dx.doi.org/10.1007/s00284-002-3960-x
---------- VANCOUVER ----------
Ruzal, S.M., Sanchez-Rivas, C. Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties. Curr. Microbiol. 2003;47(3):208-213.
http://dx.doi.org/10.1007/s00284-002-3960-x