Two-component regulatory proteins ResD-ResE are required for transcriptional activation of fnr upon oxygen limitation in Bacillus subtilis.

ABSTRACT The Bacillus subtilis sbo-alb operon containssboA, the structural gene for the bacteriocin subtilosin, and the alb genes required for subtilosin production. Transcription from the sbo-alb promoter is highly induced by oxygen limitation. The transcriptional regulation of thesbo-alb operon is under dual control involving the transition state regulator AbrB and the two-component regulatory proteins ResD and ResE.

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A Novel Class of Heat and Secretion Stress-Responsive Genes Is Controlled by the Autoregulated CssRS Two-Component System of Bacillus subtilis

Journal of Bacteriology ◽

10.1128/jb.184.20.5661-5671.2002 ◽

2002 ◽

Vol 184 (20) ◽

pp. 5661-5671 ◽

Cited By ~ 113

Author(s):

Elise Darmon ◽

David Noone ◽

Anne Masson ◽

Sierd Bron ◽

Oscar P. Kuipers ◽

...

Keyword(s):

Bacillus Subtilis ◽

Heat Stress ◽

Transcriptional Activation ◽

Cellular Response ◽

Regulatory Region ◽

Regulatory System ◽

Secretion Stress ◽

Two Component ◽

High Level ◽

Inducible Genes

ABSTRACT Bacteria need dedicated systems that allow appropriate adaptation to the perpetual changes in their environments. In Bacillus subtilis, two HtrA-like proteases, HtrA and HtrB, play critical roles in the cellular response to secretion and heat stresses. Transcription of these genes is induced by the high-level production of a secreted protein or by a temperature upshift. The CssR-CssS two-component regulatory system plays an essential role in this transcriptional activation. Transcription of the cssRS operon is autoregulated and can be induced by secretion stress, by the absence of either HtrA or HtrB, and by heat stress in a HtrA null mutant strain. Two start sites are used for cssRS transcription, only one of which is responsive to heat and secretion stress. The divergently transcribed htrB and cssRS genes share a regulatory region through which their secretion and heat stress-induced expression is linked. This study shows that CssRS-regulated genes represent a novel class of heat-inducible genes, which is referred to as class V and currently includes two genes: htrA and htrB.

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Mutational Analysis of the Signal-Sensing Domain of ResE Histidine Kinase from Bacillus subtilis

Journal of Bacteriology ◽

10.1128/jb.186.6.1694-1704.2004 ◽

2004 ◽

Vol 186 (6) ◽

pp. 1694-1704 ◽

Cited By ~ 17

Author(s):

Avanti Baruah ◽

Brett Lindsey ◽

Yi Zhu ◽

Michiko M. Nakano

Keyword(s):

Bacillus Subtilis ◽

Transcriptional Activation ◽

Catalytic Domain ◽

Mutational Analysis ◽

Response Regulator ◽

Regulatory Region ◽

Regulatory System ◽

Oxygen Limitation ◽

Nitrate Respiration ◽

Terminal Domain

ABSTRACT The Bacillus subtilis ResD-ResE two-component regulatory system activates genes involved in nitrate respiration in response to oxygen limitation or nitric oxide (NO). The sensor kinase ResE activates the response regulator ResD through phosphorylation, which then binds to the regulatory region of genes involved in anaerobiosis to activate their transcription. ResE is composed of an N-terminal signal input domain and a C-terminal catalytic domain. The N-terminal domain contains two transmembrane subdomains and a large extracytoplasmic loop. It also has a cytoplasmic PAS subdomain between the HAMP linker and C-terminal kinase domain. In an attempt to identify the signal-sensing subdomain of ResE, a series of deletions and amino acid substitutions were generated in the N-terminal domain. The results indicated that cytoplasmic ResE lacking the transmembrane segments and the extracytoplasmic loop retains the ability to sense oxygen limitation and NO, which leads to transcriptional activation of ResDE-dependent genes. This activity was eliminated by the deletion of the PAS subdomain, demonstrating that the PAS subdomain participates in signal reception. The study also raised the possibility that the extracytoplasmic region may serve as a second signal-sensing subdomain. This suggests that the extracytoplasmic region could contribute to amplification of ResE activity leading to the robust activation of genes required for anaerobic metabolism in B. subtilis.

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Transcriptional Activation by Bacillus subtilis ResD: Tandem Binding to Target Elements and Phosphorylation-Dependent and -Independent Transcriptional Activation

Journal of Bacteriology ◽

10.1128/jb.186.7.2028-2037.2004 ◽

2004 ◽

Vol 186 (7) ◽

pp. 2028-2037 ◽

Cited By ~ 21

Author(s):

Hao Geng ◽

Shunji Nakano ◽

Michiko M. Nakano

Keyword(s):

Bacillus Subtilis ◽

Transcriptional Activation ◽

Response Regulator ◽

Gene Activation ◽

Phosphorylation Site ◽

Oxygen Limitation ◽

Nitrate Respiration ◽

Expression Of Genes

ABSTRACT The expression of genes involved in nitrate respiration in Bacillus subtilis is regulated by the ResD-ResE two-component signal transduction system. The membrane-bound ResE sensor kinase perceives a redox-related signal(s) and phosphorylates the cognate response regulator ResD, which enables interaction of ResD with ResD-dependent promoters to activate transcription. Hydroxyl radical footprinting analysis revealed that ResD tandemly binds to the −41 to −83 region of hmp and the −46 to −92 region of nasD. In vitro runoff transcription experiments showed that ResD is necessary and sufficient to activate transcription of the ResDE regulon. Although phosphorylation of ResD by ResE kinase greatly stimulated transcription, unphosphorylated ResD, as well as ResD with a phosphorylation site (Asp57) mutation, was able to activate transcription at a low level. The D57A mutant was shown to retain the activity in vivo to induce transcription of the ResDE regulon in response to oxygen limitation, suggesting that ResD itself, in addition to its activation through phosphorylation-mediated conformation change, senses oxygen limitation via an unknown mechanism leading to anaerobic gene activation.

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Bacillus subtilis ResD Induces Expression of the Potential Regulatory Genes yclJK upon Oxygen Limitation

Journal of Bacteriology ◽

10.1128/jb.186.19.6477-6484.2004 ◽

2004 ◽

Vol 186 (19) ◽

pp. 6477-6484 ◽

Cited By ~ 24

Author(s):

Elisabeth Härtig ◽

Hao Geng ◽

Anja Hartmann ◽

Angela Hubacek ◽

Richard Münch ◽

...

Keyword(s):

Bacillus Subtilis ◽

Transcriptional Start Site ◽

Regulatory Region ◽

Regulatory System ◽

Oxygen Limitation ◽

Anaerobic Growth ◽

Nucleotide Substitutions ◽

Anaerobic Induction ◽

Two Component

ABSTRACT Transcription of the yclJK operon, which encodes a potential two-component regulatory system, is activated in response to oxygen limitation in Bacillus subtilis. Northern blot analysis and assays of yclJ-lacZ reporter gene fusion activity revealed that the anaerobic induction is dependent on another two-component signal transduction system encoded by resDE. ResDE was previously shown to be required for the induction of anaerobic energy metabolism. Electrophoretic mobility shift assays and DNase I footprinting experiments showed that the response regulator ResD binds specifically to the yclJK regulatory region upstream of the transcriptional start site. In vitro transcription experiments demonstrated that ResD is sufficient to activate yclJ transcription. The phosphorylation of ResD by its sensor kinase, ResE, highly stimulates its activity as a transcriptional activator. Multiple nucleotide substitutions in the ResD binding regions of the yclJ promoter abolished ResD binding in vitro and prevented the anaerobic induction of yclJK in vivo. A weight matrix for the ResD binding site was defined by a bioinformatic approach. The results obtained suggest the existence of a new branch of the complex regulatory system employed for the adaptation of B. subtilis to anaerobic growth conditions.

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Host recognition by the VirA, VirG two-component regulatory proteins of Agrobacterium tumefaciens

Research in Microbiology ◽

10.1016/0923-2508(94)90095-7 ◽

1994 ◽

Vol 145 (5-6) ◽

pp. 461-473 ◽

Cited By ~ 26

Author(s):

S.C. Winans ◽

N.J. Mantis ◽

Chin-Yi Chen ◽

Chia-Hwa Chang ◽

Dong Cho Han

Keyword(s):

Agrobacterium Tumefaciens ◽

Regulatory Proteins ◽

Host Recognition ◽

Two Component

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The Bacillus subtilis ywkA gene encodes a malic enzyme and its transcription is activated by the YufL/YufM two-component system in response to malate

Microbiology ◽

10.1099/mic.0.26256-0 ◽

2003 ◽

Vol 149 (9) ◽

pp. 2331-2343 ◽

Cited By ~ 39

Author(s):

Thierry Doan ◽

Pascale Servant ◽

Shigeo Tojo ◽

Hirotake Yamaguchi ◽

Guillaume Lerondel ◽

...

Keyword(s):

Bacillus Subtilis ◽

Malic Enzyme ◽

Northern Analysis ◽

Two Component System ◽

Extracellular Medium ◽

Component System ◽

Two Component ◽

Transcriptome Comparison ◽

Physiological Tests

A transcriptome comparison of a wild-type Bacillus subtilis strain growing under glycolytic or gluconeogenic conditions was performed. In particular, it revealed that the ywkA gene, one of the four paralogues putatively encoding a malic enzyme, was more transcribed during gluconeogenesis. Using a lacZ reporter fusion to the ywkA promoter, it was shown that ywkA was specifically induced by external malate and not subject to glucose catabolite repression. Northern analysis confirmed this expression pattern and demonstrated that ywkA is cotranscribed with the downstream ywkB gene. The ywkA gene product was purified and biochemical studies demonstrated its malic enzyme activity, which was 10-fold higher with NAD than with NADP (k cat/K m 102 and 10 s−1 mM−1, respectively). However, physiological tests with single and multiple mutant strains affected in ywkA and/or in ywkA paralogues showed that ywkA does not contribute to efficient utilization of malate for growth. Transposon mutagenesis allowed the identification of the uncharacterized YufL/YufM two-component system as being responsible for the control of ywkA expression. Genetic analysis and in vitro studies with purified YufM protein showed that YufM binds just upstream of ywkA promoter and activates ywkA transcription in response to the presence of malate in the extracellular medium, transmitted by YufL. ywkA and yufL/yufM could thus be renamed maeA for malic enzyme and malK/malR for malate kinase sensor/malate response regulator, respectively.

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