High salt stress in Bacillus subtilis: involvement of PBP4* as a peptidoglycan hydrolase

The study was focused on the role of the penicillin binding protein PBP4* of Bacillus subtilis during growth in high salinity rich media. Using pbpE-lacZ fusion, we found that transcription of the pbpE gene is induced in stationary phase and by increased salinity. This increase was also corroborated...

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Autores principales:	Palomino, María Mercedes, Ruzal, Sandra Mónica
Publicado:	2009
Materias:	Bacillus subtilis High salt Muramidase pbpE Penicillin binding protein (PBP) Peptidoglycan bacitracin beta galactosidase penicillin binding protein penicillin G peptidoglycan protein pbp 4 unclassified drug vancomycin antibiotic sensitivity article autolysis bacterial cell wall gene gene disruption gene mutation genetic transcription nonhuman operon pbpe gene priority journal racx gene salinity salt stress zymography Anti-Bacterial Agents Bacitracin Bacteriolysis Cell Wall Microscopy, Electron, Transmission N-Acetylmuramoyl-L-alanine Amidase Penicillin G Penicillin-Binding Proteins Salinity Serine-Type D-Ala-D-Ala Carboxypeptidase Transcription, Genetic Vancomycin
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09232508_v160_n2_p117_Palomino http://hdl.handle.net/20.500.12110/paper_09232508_v160_n2_p117_Palomino
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_09232508_v160_n2_p117_Palomino
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spelling	paper:paper_09232508_v160_n2_p117_Palomino2023-06-08T15:50:58Z High salt stress in Bacillus subtilis: involvement of PBP4* as a peptidoglycan hydrolase Palomino, María Mercedes Ruzal, Sandra Mónica Bacillus subtilis High salt Muramidase pbpE Penicillin binding protein (PBP) Peptidoglycan bacitracin beta galactosidase penicillin binding protein penicillin G peptidoglycan protein pbp 4 unclassified drug vancomycin antibiotic sensitivity article autolysis Bacillus subtilis bacterial cell wall gene gene disruption gene mutation genetic transcription nonhuman operon pbpe gene priority journal racx gene salinity salt stress zymography Anti-Bacterial Agents Bacillus subtilis Bacitracin Bacteriolysis Cell Wall Microscopy, Electron, Transmission N-Acetylmuramoyl-L-alanine Amidase Penicillin G Penicillin-Binding Proteins Peptidoglycan Salinity Serine-Type D-Ala-D-Ala Carboxypeptidase Transcription, Genetic Vancomycin Bacillus subtilis The study was focused on the role of the penicillin binding protein PBP4* of Bacillus subtilis during growth in high salinity rich media. Using pbpE-lacZ fusion, we found that transcription of the pbpE gene is induced in stationary phase and by increased salinity. This increase was also corroborated at the translation level for PBP4* by western blot. Furthermore, we showed that a strain harboring gene disruption in the structural gene (pbpE) for the PBP4* endopeptidase resulted in a salt-sensitive phenotype and increased sensitivity to cell envelope active antibiotics (vancomycin, penicillin and bacitracin). Since the pbpE gene seems to be part of a two-gene operon with racX, a racX::pRV300 mutant was obtained. This mutant behaved like the wild-type strain with respect to high salt. Electron microscopy showed that high salt and mutation of pbpE resulted in cell wall defects. Whole cells or purified peptidoglycan from WT cultures grown in high salt medium showed increased autolysis and susceptibility to mutanolysin. We demonstrate through zymogram analysis that PBP4* has murein hydrolyze activity. All these results support the hypothesis that peptidoglycan is modified in response to high salt and that PBP4* contributes to this modification. © 2008 Elsevier Masson SAS. All rights reserved. Fil:Palomino, M.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Ruzal, S.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2009 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09232508_v160_n2_p117_Palomino http://hdl.handle.net/20.500.12110/paper_09232508_v160_n2_p117_Palomino
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Bacillus subtilis High salt Muramidase pbpE Penicillin binding protein (PBP) Peptidoglycan bacitracin beta galactosidase penicillin binding protein penicillin G peptidoglycan protein pbp 4 unclassified drug vancomycin antibiotic sensitivity article autolysis Bacillus subtilis bacterial cell wall gene gene disruption gene mutation genetic transcription nonhuman operon pbpe gene priority journal racx gene salinity salt stress zymography Anti-Bacterial Agents Bacillus subtilis Bacitracin Bacteriolysis Cell Wall Microscopy, Electron, Transmission N-Acetylmuramoyl-L-alanine Amidase Penicillin G Penicillin-Binding Proteins Peptidoglycan Salinity Serine-Type D-Ala-D-Ala Carboxypeptidase Transcription, Genetic Vancomycin Bacillus subtilis
spellingShingle	Bacillus subtilis High salt Muramidase pbpE Penicillin binding protein (PBP) Peptidoglycan bacitracin beta galactosidase penicillin binding protein penicillin G peptidoglycan protein pbp 4 unclassified drug vancomycin antibiotic sensitivity article autolysis Bacillus subtilis bacterial cell wall gene gene disruption gene mutation genetic transcription nonhuman operon pbpe gene priority journal racx gene salinity salt stress zymography Anti-Bacterial Agents Bacillus subtilis Bacitracin Bacteriolysis Cell Wall Microscopy, Electron, Transmission N-Acetylmuramoyl-L-alanine Amidase Penicillin G Penicillin-Binding Proteins Peptidoglycan Salinity Serine-Type D-Ala-D-Ala Carboxypeptidase Transcription, Genetic Vancomycin Bacillus subtilis Palomino, María Mercedes Ruzal, Sandra Mónica High salt stress in Bacillus subtilis: involvement of PBP4* as a peptidoglycan hydrolase
topic_facet	Bacillus subtilis High salt Muramidase pbpE Penicillin binding protein (PBP) Peptidoglycan bacitracin beta galactosidase penicillin binding protein penicillin G peptidoglycan protein pbp 4 unclassified drug vancomycin antibiotic sensitivity article autolysis Bacillus subtilis bacterial cell wall gene gene disruption gene mutation genetic transcription nonhuman operon pbpe gene priority journal racx gene salinity salt stress zymography Anti-Bacterial Agents Bacillus subtilis Bacitracin Bacteriolysis Cell Wall Microscopy, Electron, Transmission N-Acetylmuramoyl-L-alanine Amidase Penicillin G Penicillin-Binding Proteins Peptidoglycan Salinity Serine-Type D-Ala-D-Ala Carboxypeptidase Transcription, Genetic Vancomycin Bacillus subtilis
description	The study was focused on the role of the penicillin binding protein PBP4* of Bacillus subtilis during growth in high salinity rich media. Using pbpE-lacZ fusion, we found that transcription of the pbpE gene is induced in stationary phase and by increased salinity. This increase was also corroborated at the translation level for PBP4* by western blot. Furthermore, we showed that a strain harboring gene disruption in the structural gene (pbpE) for the PBP4* endopeptidase resulted in a salt-sensitive phenotype and increased sensitivity to cell envelope active antibiotics (vancomycin, penicillin and bacitracin). Since the pbpE gene seems to be part of a two-gene operon with racX, a racX::pRV300 mutant was obtained. This mutant behaved like the wild-type strain with respect to high salt. Electron microscopy showed that high salt and mutation of pbpE resulted in cell wall defects. Whole cells or purified peptidoglycan from WT cultures grown in high salt medium showed increased autolysis and susceptibility to mutanolysin. We demonstrate through zymogram analysis that PBP4* has murein hydrolyze activity. All these results support the hypothesis that peptidoglycan is modified in response to high salt and that PBP4* contributes to this modification. © 2008 Elsevier Masson SAS. All rights reserved.
author	Palomino, María Mercedes Ruzal, Sandra Mónica
author_facet	Palomino, María Mercedes Ruzal, Sandra Mónica
author_sort	Palomino, María Mercedes
title	High salt stress in Bacillus subtilis: involvement of PBP4* as a peptidoglycan hydrolase
title_short	High salt stress in Bacillus subtilis: involvement of PBP4* as a peptidoglycan hydrolase
title_full	High salt stress in Bacillus subtilis: involvement of PBP4* as a peptidoglycan hydrolase
title_fullStr	High salt stress in Bacillus subtilis: involvement of PBP4* as a peptidoglycan hydrolase
title_full_unstemmed	High salt stress in Bacillus subtilis: involvement of PBP4* as a peptidoglycan hydrolase
title_sort	high salt stress in bacillus subtilis: involvement of pbp4* as a peptidoglycan hydrolase
publishDate	2009
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09232508_v160_n2_p117_Palomino http://hdl.handle.net/20.500.12110/paper_09232508_v160_n2_p117_Palomino
work_keys_str_mv	AT palominomariamercedes highsaltstressinbacillussubtilisinvolvementofpbp4asapeptidoglycanhydrolase AT ruzalsandramonica highsaltstressinbacillussubtilisinvolvementofpbp4asapeptidoglycanhydrolase
_version_	1768542847257018368

High salt stress in Bacillus subtilis: involvement of PBP4* as a peptidoglycan hydrolase

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