Role of serine/threonine protein phosphatase PrpN in the life cycle of Bacillus anthracis
Reversible protein phosphorylation at serine/threonine residues is one of the most common protein modifications, widely observed in all kingdoms of life. The catalysts controlling this modification are specific serine/threonine kinases and phosphatases that modulate various cellular pathways ranging...
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Published in | PLoS pathogens Vol. 18; no. 8; p. e1010729 |
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Main Authors | , , , , , , , , , , |
Format | Journal Article |
Language | English |
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Public Library of Science
01.08.2022
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Abstract | Reversible protein phosphorylation at serine/threonine residues is one of the most common protein modifications, widely observed in all kingdoms of life. The catalysts controlling this modification are specific serine/threonine kinases and phosphatases that modulate various cellular pathways ranging from growth to cellular death. Genome sequencing and various omics studies have led to the identification of numerous serine/threonine kinases and cognate phosphatases, yet the physiological relevance of many of these proteins remain enigmatic. In
Bacillus anthracis
, only one ser/thr phosphatase, PrpC, has been functionally characterized; it was reported to be non-essential for bacterial growth and survival. In the present study, we characterized another ser/thr phosphatase (PrpN) of
B
.
anthracis
by various structural and functional approaches. To examine its physiological relevance in
B
.
anthracis
, a null mutant strain of
prpN
was generated and shown to have defects in sporulation and reduced synthesis of toxins (PA and LF) and the toxin activator protein AtxA. We also identified CodY, a global transcriptional regulator, as a target of PrpN and ser/thr kinase PrkC. CodY phosphorylation strongly controlled its binding to the promoter region of
atxA
, as shown using phosphomimetic and phosphoablative mutants. In nutshell, the present study reports phosphorylation-mediated regulation of CodY activity in the context of anthrax toxin synthesis in
B
.
anthracis
by a previously uncharacterized ser/thr protein phosphatase–PrpN. |
---|---|
AbstractList | Reversible protein phosphorylation at serine/threonine residues is one of the most common protein modifications, widely observed in all kingdoms of life. The catalysts controlling this modification are specific serine/threonine kinases and phosphatases that modulate various cellular pathways ranging from growth to cellular death. Genome sequencing and various omics studies have led to the identification of numerous serine/threonine kinases and cognate phosphatases, yet the physiological relevance of many of these proteins remain enigmatic. In
Bacillus anthracis
, only one ser/thr phosphatase, PrpC, has been functionally characterized; it was reported to be non-essential for bacterial growth and survival. In the present study, we characterized another ser/thr phosphatase (PrpN) of
B
.
anthracis
by various structural and functional approaches. To examine its physiological relevance in
B
.
anthracis
, a null mutant strain of
prpN
was generated and shown to have defects in sporulation and reduced synthesis of toxins (PA and LF) and the toxin activator protein AtxA. We also identified CodY, a global transcriptional regulator, as a target of PrpN and ser/thr kinase PrkC. CodY phosphorylation strongly controlled its binding to the promoter region of
atxA
, as shown using phosphomimetic and phosphoablative mutants. In nutshell, the present study reports phosphorylation-mediated regulation of CodY activity in the context of anthrax toxin synthesis in
B
.
anthracis
by a previously uncharacterized ser/thr protein phosphatase–PrpN. Reversible protein phosphorylation at serine/threonine residues is one of the most common protein modifications, widely observed in all kingdoms of life. The catalysts controlling this modification are specific serine/threonine kinases and phosphatases that modulate various cellular pathways ranging from growth to cellular death. Genome sequencing and various omics studies have led to the identification of numerous serine/threonine kinases and cognate phosphatases, yet the physiological relevance of many of these proteins remain enigmatic. In Bacillus anthracis, only one ser/thr phosphatase, PrpC, has been functionally characterized; it was reported to be non-essential for bacterial growth and survival. In the present study, we characterized another ser/thr phosphatase (PrpN) of B. anthracis by various structural and functional approaches. To examine its physiological relevance in B. anthracis, a null mutant strain of prpN was generated and shown to have defects in sporulation and reduced synthesis of toxins (PA and LF) and the toxin activator protein AtxA. We also identified CodY, a global transcriptional regulator, as a target of PrpN and ser/thr kinase PrkC. CodY phosphorylation strongly controlled its binding to the promoter region of atxA, as shown using phosphomimetic and phosphoablative mutants. In nutshell, the present study reports phosphorylation-mediated regulation of CodY activity in the context of anthrax toxin synthesis in B. anthracis by a previously uncharacterized ser/thr protein phosphatase–PrpN. Reversible protein phosphorylation at serine/threonine residues is one of the most common protein modifications, widely observed in all kingdoms of life. The catalysts controlling this modification are specific serine/threonine kinases and phosphatases that modulate various cellular pathways ranging from growth to cellular death. Genome sequencing and various omics studies have led to the identification of numerous serine/threonine kinases and cognate phosphatases, yet the physiological relevance of many of these proteins remain enigmatic. In Bacillus anthracis , only one ser/thr phosphatase, PrpC, has been functionally characterized; it was reported to be non-essential for bacterial growth and survival. In the present study, we characterized another ser/thr phosphatase (PrpN) of B . anthracis by various structural and functional approaches. To examine its physiological relevance in B . anthracis , a null mutant strain of prpN was generated and shown to have defects in sporulation and reduced synthesis of toxins (PA and LF) and the toxin activator protein AtxA. We also identified CodY, a global transcriptional regulator, as a target of PrpN and ser/thr kinase PrkC. CodY phosphorylation strongly controlled its binding to the promoter region of atxA , as shown using phosphomimetic and phosphoablative mutants. In nutshell, the present study reports phosphorylation-mediated regulation of CodY activity in the context of anthrax toxin synthesis in B . anthracis by a previously uncharacterized ser/thr protein phosphatase–PrpN. Reversible protein phosphorylation at specific ser/thr residues causes conformational changes in the protein structure, thereby modulating its cellular activity. In B . anthracis , though the role of ser/thr phosphorylation is implicated in various cellular pathways including pathogenesis, till date only one STP (PrpC) has been functionally characterized. This manuscript reports functional characterization of another STP (PrpN) in B . anthracis and with the aid of a null mutant strain (BAS Δ prpN ) we provide important insight regarding the role of PrpN in the life cycle of B . anthracis . We have also identified the global transcriptional regulator, CodY as a target of PrpN and PrkC, and for the first time showed the physiological relevance of CodY phosphorylation status in the regulation of anthrax toxin synthesis. |
Audience | Academic |
Author | Singh, Lalit K Bothra, Ankur Leppla, Stephen H Kumar, Nishant Gangwal, Aakriti Sangwan, Nitika Keshavam, Chetkar Chandra Pomerantsev, Andrei P Dhasmana, Neha Goel, Ajay K Singh, Yogendra |
AuthorAffiliation | 4 Division of Biotechnology, Defence Research and Development Establishment, Gwalior, Madhya Pradesh, India 1 Department of Zoology, University of Delhi, Delhi, India 3 Microbial Pathogenesis Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, United States of America 2 CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi, India University of Texas Medical School at Houston, UNITED STATES |
AuthorAffiliation_xml | – name: 2 CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi, India – name: 1 Department of Zoology, University of Delhi, Delhi, India – name: 4 Division of Biotechnology, Defence Research and Development Establishment, Gwalior, Madhya Pradesh, India – name: 3 Microbial Pathogenesis Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, United States of America – name: University of Texas Medical School at Houston, UNITED STATES |
Author_xml | – sequence: 1 fullname: Gangwal, Aakriti – sequence: 2 fullname: Sangwan, Nitika – sequence: 3 fullname: Dhasmana, Neha – sequence: 4 fullname: Kumar, Nishant – sequence: 5 fullname: Keshavam, Chetkar Chandra – sequence: 6 fullname: Singh, Lalit K – sequence: 7 fullname: Bothra, Ankur – sequence: 8 fullname: Goel, Ajay K – sequence: 9 fullname: Pomerantsev, Andrei P – sequence: 10 fullname: Leppla, Stephen H – sequence: 11 fullname: Singh, Yogendra |
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CitedBy_id | crossref_primary_10_1093_femsre_fuad044 crossref_primary_10_1111_mmi_15220 |
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Notes | new_version ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 The authors have declared that no competing interests exist. Current address: InVitaGO Diagnostic GmbH, Frankfurt am Main, Germany. |
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SubjectTerms | Anthrax Bacillus anthracis Bacterial proteins Binding sites Biology and Life Sciences Catalysts Cell death Chemical properties Edema Enzyme kinetics Gene sequencing Genes Genomes Glycerol Growth Kinases Life cycles Medicine and Health Sciences Mutants Pathogens Phosphatase Phosphatases Phosphorylation Physiological aspects Physiology Plasmids Prokaryotes Protein kinases Protein phosphatase Protein-serine/threonine kinase Proteins Regulation Serine Sporulation Streptococcus infections Structure-function relationships Synthesis Threonine Toxins Virulence |
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Title | Role of serine/threonine protein phosphatase PrpN in the life cycle of Bacillus anthracis |
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