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 inPLoS pathogens Vol. 18; no. 8; p. e1010729
Main Authors Gangwal, Aakriti, Sangwan, Nitika, Dhasmana, Neha, Kumar, Nishant, Keshavam, Chetkar Chandra, Singh, Lalit K, Bothra, Ankur, Goel, Ajay K, Pomerantsev, Andrei P, Leppla, Stephen H, Singh, Yogendra
Format Journal Article
LanguageEnglish
Published San Francisco 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
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– name: 1 Department of Zoology, University of Delhi, Delhi, India
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– name: University of Texas Medical School at Houston, UNITED STATES
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The authors have declared that no competing interests exist.
Current address: InVitaGO Diagnostic GmbH, Frankfurt am Main, Germany.
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SSID ssj0041316
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Snippet Reversible protein phosphorylation at serine/threonine residues is one of the most common protein modifications, widely observed in all kingdoms of life. The...
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StartPage e1010729
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
URI https://www.proquest.com/docview/2715142498
https://search.proquest.com/docview/2697092077
https://pubmed.ncbi.nlm.nih.gov/PMC9371265
https://doaj.org/article/9fd5a241a92149bda4fd11b8827b77eb
http://dx.doi.org/10.1371/journal.ppat.1010729
Volume 18
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