Die Zellwand-Hydrolase YocH aus Bacillus subtilis: Genetische Kontrolle durch das essentielle Zwei-Komponenten System YycFG, hohe Osmolarität und Kältestress.

Bakterien besiedeln die unterschiedlichsten Lebensräume. Diese Habitate unterliegen oftmals großen Schwankungen biotischer und abiotischer Faktoren, denen die bakterielle Zelle ausgeliefert ist und auf die sie zeitgerecht reagieren muss, um ihr Wachstum und Überleben zu sichern. Das Habitat des Gram...

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1. Verfasser: Seibert, Tim Martin
Beteiligte: Bremer, Erhard (Prof. Dr.) (BetreuerIn (Doktorarbeit))
Format: Dissertation
Sprache:Deutsch
Veröffentlicht: Philipps-Universität Marburg 2009
Biologie
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topic two-component system YycFG
osmotic stress
Zwei-Komponenten System YycFG
cell wall hydrolase
Kältestress
Biowissenschaften, Biologie
Zellwand-Hydrolase
Osmotischer Stress
Heubacillus
Zellwand
Murein
cold stress
spellingShingle two-component system YycFG
osmotic stress
Zwei-Komponenten System YycFG
cell wall hydrolase
Kältestress
Biowissenschaften, Biologie
Zellwand-Hydrolase
Osmotischer Stress
Heubacillus
Zellwand
Murein
cold stress
Bacteria are able to colonize a large quantity of different environments. These habitats underlie extensive variations of biotic and abiotic factors to which the bacterial cell has to respond timely to ensure growth and survival. The habitat of the gram-positive bacterium Bacillus subtilis is the upper layers of the soil and the rizosphere. Here two of the most important abiotic factors are directly influencing the bacterial cell. B. subtilis is due to alteration of day and night, weather conditions and changing seasons, exposed to constant variations in osmolarity and temperature. DNA-array analysis of B. subtilis have shown, that there are a lot of genes involved in cell wall metabolism which are induced by hyperosmotic conditions and adaptive growth at 15°C (Steil et al., 2003, Budde et al., 2006). This is a hint, that the proteins coded by these genes play important roles in the adaptation of cell wall structure and composition of B. subtilis under stress conditions. In regard to that, it could be shown before for several bacterial species that the cell envelope and especially the cell wall is changing upon increase in osmolarity and a drop in temperature (Vijaranakul et al., 1995, Lopez et al., 1998, Lopez et al., 2000, Piuri et al., 2005, Palomino et al., 2008). One of the genes which could be identified as induced by osmotic- and cold stress in B. subtilis is the yocH gene. In the present dissertation the genetic regulation of the yocH gene and the function of the YocH protein could be characterised in detail. Due to biochemical analysis of the purified YocH protein and an analysis of a YocH-GFP fusion it could be shown, that YocH is a peptidoglycan associated cell wall hydrolase. YocH has an important role in the dynamic reconstruction of the peptidoglycan during growth under stress condtions, as a yocH deletion mutant is sensitive to high osmolarity. The expression of the yocH gene is controlled be the single essential two-component regulatory system (YycFG) of B. subtilis (Howell et al., 2003; Dubrac et al., 2008). The presented data gain a first insight into the architecture of the regulatory region of yocH and the transcriptional regulation of this gene responding to hyperosmotic conditions and a drop in growth temperature. The importance of the YycFG system and the transition-state regulator AbrB onto the induction of the yocH promoter under hyperosmotic condtions and at low temperature (15°C) could be shown in detail.
Seibert, Tim Martin
Die Zellwand-Hydrolase YocH aus Bacillus subtilis: Genetische Kontrolle durch das essentielle Zwei-Komponenten System YycFG, hohe Osmolarität und Kältestress.
contents Bacteria are able to colonize a large quantity of different environments. These habitats underlie extensive variations of biotic and abiotic factors to which the bacterial cell has to respond timely to ensure growth and survival. The habitat of the gram-positive bacterium Bacillus subtilis is the upper layers of the soil and the rizosphere. Here two of the most important abiotic factors are directly influencing the bacterial cell. B. subtilis is due to alteration of day and night, weather conditions and changing seasons, exposed to constant variations in osmolarity and temperature. DNA-array analysis of B. subtilis have shown, that there are a lot of genes involved in cell wall metabolism which are induced by hyperosmotic conditions and adaptive growth at 15°C (Steil et al., 2003, Budde et al., 2006). This is a hint, that the proteins coded by these genes play important roles in the adaptation of cell wall structure and composition of B. subtilis under stress conditions. In regard to that, it could be shown before for several bacterial species that the cell envelope and especially the cell wall is changing upon increase in osmolarity and a drop in temperature (Vijaranakul et al., 1995, Lopez et al., 1998, Lopez et al., 2000, Piuri et al., 2005, Palomino et al., 2008). One of the genes which could be identified as induced by osmotic- and cold stress in B. subtilis is the yocH gene. In the present dissertation the genetic regulation of the yocH gene and the function of the YocH protein could be characterised in detail. Due to biochemical analysis of the purified YocH protein and an analysis of a YocH-GFP fusion it could be shown, that YocH is a peptidoglycan associated cell wall hydrolase. YocH has an important role in the dynamic reconstruction of the peptidoglycan during growth under stress condtions, as a yocH deletion mutant is sensitive to high osmolarity. The expression of the yocH gene is controlled be the single essential two-component regulatory system (YycFG) of B. subtilis (Howell et al., 2003; Dubrac et al., 2008). The presented data gain a first insight into the architecture of the regulatory region of yocH and the transcriptional regulation of this gene responding to hyperosmotic conditions and a drop in growth temperature. The importance of the YycFG system and the transition-state regulator AbrB onto the induction of the yocH promoter under hyperosmotic condtions and at low temperature (15°C) could be shown in detail.
publishDate 2009
era_facet 2009
language German
author2 Bremer, Erhard (Prof. Dr.)
author2_role ths
format Dissertation
oai_set_str_mv doc-type:doctoralThesis
ddc:570
open_access
xMetaDissPlus
description Bakterien besiedeln die unterschiedlichsten Lebensräume. Diese Habitate unterliegen oftmals großen Schwankungen biotischer und abiotischer Faktoren, denen die bakterielle Zelle ausgeliefert ist und auf die sie zeitgerecht reagieren muss, um ihr Wachstum und Überleben zu sichern. Das Habitat des Gram-positiven Bakteriums Bacillus subtilis sind die oberen Bodenschichten und die Rizosphäre. Hier nehmen zwei der wichtigsten abiotischen Wachstumsfaktoren unmittelbaren Einfluss auf die bakterielle Zelle. So ist B. subtilis durch Tag- und Nachtwechsel, Wetteränderungen und jahreszeitliche Unterschiede ständigen Schwankungen in Osmolarität und Temperatur unterworfen. DNA-Array Analysen bei B. subtilis haben gezeigt, dass eine Reihe von Genen, die im Zusammenhang mit dem Zellwandmetabolismus stehen durch hyperosmotische Bedingungen und adaptives Wachstum bei 15°C induziert werden (Steil et al., 2003, Budde et al., 2006). Dies deutet darauf hin, dass die von diesen Genen kodierten Proteine wesentliche Funktionen für die Anpassung der Zellwand-Struktur und -Zusammensetzung von B. subtilis unter Stress-Bediungungen ausüben. In diesem Zusammenhang konnten zuvor schon Veränderungen der Zellhülle - und insbesondere der Zellwand - als Antwort auf erhöhte Osmolarität und sinkende Wachstumstemperaturen bei verschiedenen Species nachgewiesen werden (Vijaranakul et al., 1995, Lopez et al., 1998, Lopez et al., 2000, Piuri et al., 2005, Palomino et al., 2008). Eines der durch die DNA-Arrays von osmotisch- und Kälte-gestressten B. subtilis Zellen in den Fokus des Interesses gerückten Gene ist yocH. Im Rahmen der vorliegenden Arbeit wurde die genetische Regulation des yocH Gens und die Funktion des YocH Proteins näher charakterisiert. Es konnte durch biochemische Analyse des gereinigten YocH Proteins und durch Untersuchungen mit einer YocH-GFP Fusion gezeigt werden, dass YocH als eine Peptidoglykan-assoziierte Zellwandhydrolase fungiert. YocH spielt eine wichtige Rolle beim dynamischen Umbau des Peptidoglykans während des Wachstums unter Stressbedingungen, da eine yocH Mutante osmotisch sensitiv ist. Die Expression des yocH Gens unterliegt der Kontrolle durch das einzige essentielle Zwei-Komponenten Regulationssystem (YycFG) von B. subtilis (Howell et al., 2003; Dubrac et al., 2008). Die hier vorgelegten Daten erlauben erstmals einen genaueren Einblick in die Architektur der yocH Kontrollregion und in die Regulation der Expression von yocH in Antwort auf eine Erhöhung der Osmolarität und eine Absenkung der Wachstumstemperatur. Die Bedeutung des YycFG Systems und des „transition-state“ Regulators AbrB auf die Induzierbarkeit des yocH Promotors bei adaptivem Wachstum bei hoher Osmolarität und bei niedriger Temperatur (15°C) wurde herausgearbeitet.
dewey-raw 570
dewey-search 570
genre Life sciences
genre_facet Life sciences
topic_facet Biowissenschaften, Biologie
building Fachbereich Biologie
author Seibert, Tim Martin
license_str http://archiv.ub.uni-marburg.de/adm/urhg.html
title_alt The cell wall hydrolase YocH from Bacillus subtilis: Genetic control by the essential two-component system YycFG, high osmolarity and cold stress.
url http://archiv.ub.uni-marburg.de/diss/z2010/0132/pdf/dtms.pdf
last_indexed 2011-08-10T23:59:59Z
institution Biologie
publisher Philipps-Universität Marburg
ref_str_mv references
title Die Zellwand-Hydrolase YocH aus Bacillus subtilis: Genetische Kontrolle durch das essentielle Zwei-Komponenten System YycFG, hohe Osmolarität und Kältestress.
title_short Die Zellwand-Hydrolase YocH aus Bacillus subtilis: Genetische Kontrolle durch das essentielle Zwei-Komponenten System YycFG, hohe Osmolarität und Kältestress.
title_full Die Zellwand-Hydrolase YocH aus Bacillus subtilis: Genetische Kontrolle durch das essentielle Zwei-Komponenten System YycFG, hohe Osmolarität und Kältestress.
title_fullStr Die Zellwand-Hydrolase YocH aus Bacillus subtilis: Genetische Kontrolle durch das essentielle Zwei-Komponenten System YycFG, hohe Osmolarität und Kältestress.
title_full_unstemmed Die Zellwand-Hydrolase YocH aus Bacillus subtilis: Genetische Kontrolle durch das essentielle Zwei-Komponenten System YycFG, hohe Osmolarität und Kältestress.
title_sort Die Zellwand-Hydrolase YocH aus Bacillus subtilis: Genetische Kontrolle durch das essentielle Zwei-Komponenten System YycFG, hohe Osmolarität und Kältestress.
first_indexed 2010-05-12T00:00:00Z
thumbnail http://archiv.ub.uni-marburg.de/diss/z2010/0132/cover.png
spelling diss/z2010/0132 Bacteria are able to colonize a large quantity of different environments. These habitats underlie extensive variations of biotic and abiotic factors to which the bacterial cell has to respond timely to ensure growth and survival. The habitat of the gram-positive bacterium Bacillus subtilis is the upper layers of the soil and the rizosphere. Here two of the most important abiotic factors are directly influencing the bacterial cell. B. subtilis is due to alteration of day and night, weather conditions and changing seasons, exposed to constant variations in osmolarity and temperature. DNA-array analysis of B. subtilis have shown, that there are a lot of genes involved in cell wall metabolism which are induced by hyperosmotic conditions and adaptive growth at 15°C (Steil et al., 2003, Budde et al., 2006). This is a hint, that the proteins coded by these genes play important roles in the adaptation of cell wall structure and composition of B. subtilis under stress conditions. In regard to that, it could be shown before for several bacterial species that the cell envelope and especially the cell wall is changing upon increase in osmolarity and a drop in temperature (Vijaranakul et al., 1995, Lopez et al., 1998, Lopez et al., 2000, Piuri et al., 2005, Palomino et al., 2008). One of the genes which could be identified as induced by osmotic- and cold stress in B. subtilis is the yocH gene. In the present dissertation the genetic regulation of the yocH gene and the function of the YocH protein could be characterised in detail. Due to biochemical analysis of the purified YocH protein and an analysis of a YocH-GFP fusion it could be shown, that YocH is a peptidoglycan associated cell wall hydrolase. YocH has an important role in the dynamic reconstruction of the peptidoglycan during growth under stress condtions, as a yocH deletion mutant is sensitive to high osmolarity. The expression of the yocH gene is controlled be the single essential two-component regulatory system (YycFG) of B. subtilis (Howell et al., 2003; Dubrac et al., 2008). The presented data gain a first insight into the architecture of the regulatory region of yocH and the transcriptional regulation of this gene responding to hyperosmotic conditions and a drop in growth temperature. The importance of the YycFG system and the transition-state regulator AbrB onto the induction of the yocH promoter under hyperosmotic condtions and at low temperature (15°C) could be shown in detail. 2009 urn:nbn:de:hebis:04-z2010-01327 Bakterien besiedeln die unterschiedlichsten Lebensräume. Diese Habitate unterliegen oftmals großen Schwankungen biotischer und abiotischer Faktoren, denen die bakterielle Zelle ausgeliefert ist und auf die sie zeitgerecht reagieren muss, um ihr Wachstum und Überleben zu sichern. Das Habitat des Gram-positiven Bakteriums Bacillus subtilis sind die oberen Bodenschichten und die Rizosphäre. Hier nehmen zwei der wichtigsten abiotischen Wachstumsfaktoren unmittelbaren Einfluss auf die bakterielle Zelle. So ist B. subtilis durch Tag- und Nachtwechsel, Wetteränderungen und jahreszeitliche Unterschiede ständigen Schwankungen in Osmolarität und Temperatur unterworfen. DNA-Array Analysen bei B. subtilis haben gezeigt, dass eine Reihe von Genen, die im Zusammenhang mit dem Zellwandmetabolismus stehen durch hyperosmotische Bedingungen und adaptives Wachstum bei 15°C induziert werden (Steil et al., 2003, Budde et al., 2006). Dies deutet darauf hin, dass die von diesen Genen kodierten Proteine wesentliche Funktionen für die Anpassung der Zellwand-Struktur und -Zusammensetzung von B. subtilis unter Stress-Bediungungen ausüben. In diesem Zusammenhang konnten zuvor schon Veränderungen der Zellhülle - und insbesondere der Zellwand - als Antwort auf erhöhte Osmolarität und sinkende Wachstumstemperaturen bei verschiedenen Species nachgewiesen werden (Vijaranakul et al., 1995, Lopez et al., 1998, Lopez et al., 2000, Piuri et al., 2005, Palomino et al., 2008). Eines der durch die DNA-Arrays von osmotisch- und Kälte-gestressten B. subtilis Zellen in den Fokus des Interesses gerückten Gene ist yocH. Im Rahmen der vorliegenden Arbeit wurde die genetische Regulation des yocH Gens und die Funktion des YocH Proteins näher charakterisiert. Es konnte durch biochemische Analyse des gereinigten YocH Proteins und durch Untersuchungen mit einer YocH-GFP Fusion gezeigt werden, dass YocH als eine Peptidoglykan-assoziierte Zellwandhydrolase fungiert. YocH spielt eine wichtige Rolle beim dynamischen Umbau des Peptidoglykans während des Wachstums unter Stressbedingungen, da eine yocH Mutante osmotisch sensitiv ist. Die Expression des yocH Gens unterliegt der Kontrolle durch das einzige essentielle Zwei-Komponenten Regulationssystem (YycFG) von B. subtilis (Howell et al., 2003; Dubrac et al., 2008). Die hier vorgelegten Daten erlauben erstmals einen genaueren Einblick in die Architektur der yocH Kontrollregion und in die Regulation der Expression von yocH in Antwort auf eine Erhöhung der Osmolarität und eine Absenkung der Wachstumstemperatur. Die Bedeutung des YycFG Systems und des „transition-state“ Regulators AbrB auf die Induzierbarkeit des yocH Promotors bei adaptivem Wachstum bei hoher Osmolarität und bei niedriger Temperatur (15°C) wurde herausgearbeitet. 2009-06-26 The cell wall hydrolase YocH from Bacillus subtilis: Genetic control by the essential two-component system YycFG, high osmolarity and cold stress. 2011-08-10 Laemmli, U. K., (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227: 680-5. 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Arnold, K., L. Bordoli, J. Kopp & T. Schwede, (2006) The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics 22: 195-201. 2006 The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling Zuber, B., M. Haenni, T. Ribeiro, K. Minnig, F. Lopes, P. Moreillon & J. 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