Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties

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...

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Autor principal:	Ruzal, Sandra Mónica
Publicado:	2003
Materias:	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 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 Bacteria (microorganisms) Felis catus insertion sequences Posibacteria transposons
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03438651_v47_n3_p208_Ruzal http://hdl.handle.net/20.500.12110/paper_03438651_v47_n3_p208_Ruzal
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_03438651_v47_n3_p208_Ruzal
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spelling	paper:paper_03438651_v47_n3_p208_Ruzal2023-06-08T15:34:32Z Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties Ruzal, Sandra Mónica 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 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. Fil:Ruzal, S.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2003 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03438651_v47_n3_p208_Ruzal http://hdl.handle.net/20.500.12110/paper_03438651_v47_n3_p208_Ruzal
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	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
spellingShingle	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 Ruzal, Sandra Mónica Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties
topic_facet	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
description	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.
author	Ruzal, Sandra Mónica
author_facet	Ruzal, Sandra Mónica
author_sort	Ruzal, Sandra Mónica
title	Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties
title_short	Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties
title_full	Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties
title_fullStr	Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties
title_full_unstemmed	Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties
title_sort	effect of glutamate synthase (gogat) activity on bacillus subtilis spore properties
publishDate	2003
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03438651_v47_n3_p208_Ruzal http://hdl.handle.net/20.500.12110/paper_03438651_v47_n3_p208_Ruzal
work_keys_str_mv	AT ruzalsandramonica effectofglutamatesynthasegogatactivityonbacillussubtilissporeproperties
_version_	1768545834248437760

Effect of glutamate synthase (GOGAT) activity on Bacillus subtilis spore properties

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