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Cited by 2Bacillus and Francisella bacterial researchBacterial Genetics and BiotechnologyEscherichia coli research studies

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Environmental regulation of toxin production in Bacillus anthracis

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Bothra, Ankur, Pomerantsev, Andrei, Schwarz, Benjamin, Mondal, Anupam, Bohrnsen, Eric, Sangwan, Nitika, Stromberg, Kaitlin A., Moayeri, Mahtab, Ma, Qian, Fattah, Rasem, Ganesan, Sundar, Bosio, Catharine M., Leppla, Stephen H.·2025·PLOS Pathogens
DOI 10.1371/journal.ppat.1013587PMC PMC12680359

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Abstract

Pathogenic Bacillus anthracis strains carry two plasmids - pX01, which encodes a tripartite protein exotoxin complex (PA, LF, and EF); and pX02, which encodes a poly-D-gamma-glutamic acid capsule. A multidomain transcription factor, AtxA, regulates the expression of these virulence genes. AtxA has two DNA-binding Helix-Turn-Helix (HTH) domains, two phosphoenolpyruvate: carbohydrate phosphotransferase system regulatory domains (PRD1 and PRD2), and a putative EIIB domain (a component of PTS sugar transport EII-complexes). Previous studies showed that glucose and CO 2 increase AtxA-dependent toxin gene transcription, along with histidine phosphorylation of PRD1 and PRD2. Our transcriptional profiling of virulence factors, PA secretion, and fluorescent reporter strain analyses confirms a synergistic effect of glucose and CO 2 on AtxA-dependent toxin production. Deletion of AtxA (Δ atxA ) significantly reduced glucose uptake in bacteria, suggesting that AtxA may act within the glucose-PTS system. Mutation analysis of the EIIB domain of AtxA identified the cysteine at position 402 as essential for the transcriptional activity of AtxA. Deletion of glucose PTS permease PtsG (Δ ptsG ) significantly reduced the expression of PA, LF, and EF. Loss of PtsG also caused attenuation in a mouse model of infection. Intracellular imaging using FLIM confirms a physical interaction of PtsG and AtxA through EIIB domain of AtxA. Using phosphomimetic and phosphoablative mutants of AtxA, we confirmed that the physical interaction of PtsG and AtxA is essential for AtxA activity. Finally, the synergy between glucose and CO 2 was targeted by deleting pyruvate carboxylase Pyc (Δ pyc ), which regulates anaplerosis. This deletion confirms that Pyc stimulates the level of phosphoenolpyruvate (PEP) and increases the phosphorelay in glucose-PTS to enhance AtxA activity. Therefore, we propose that a histidine-phosphorelay from PEP regulates AtxA via PTS enzymatic activity, impacting AtxA activity through physical interaction of AtxA and PtsG. Finally, we propose AtxA as an integral component of the glucose-PTS, where transcriptional activity of AtxA is regulated by environmental signals including glucose and CO 2 .

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Metadata

Authors
Bothra, A., Pomerantsev, A., Schwarz, B., Mondal, A., Bohrnsen, E., Sangwan, N., Stromberg, K. A., Moayeri, M., Ma, Q., Fattah, R., Ganesan, S., Bosio, C. M., & Leppla, S. H.
Affiliations
National Institutes of Health (US)
Corresponding author
Bothra, Ankur
ORCID
Venue
PLOS Pathogens
Publisher
Public Library of Science (PLoS)
Published
2025-12-01
Volume
21
Issue
12
Pages
e1013587
ISSN
  • 1553-7366 (ISSN-L)
  • 1553-7374
DOI
10.1371/journal.ppat.1013587
PMC
PMC12680359
OpenAlex
W4416946995
Cited by
2
References
2
Citations by year
2026: 1 · 2025: 1
FWCI
0.60
Type
journal article
Language
en
Update notices
new_version (2025-12-05)
Open access
Licence
Locations
  • PLoS Pathogens (publishedVersion, public-domain)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, public-domain)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, public-domain)
Funders
Division of Intramural Research, National Institute of Allergy and Infectious Diseases
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bacillus and Francisella bacterial research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Immunology and Microbiology › Microbiology › Escherichia coli research studies
Keywords
Bacillus anthracis, AtxA, toxin gene regulation, pyruvate carboxylase, anaplerosis, carbon dioxide, histidine phosphorylation, virulence gene expression, protein-protein interaction
MeSH terms
Animals, Anthrax, Antigens, Bacterial, Bacillus anthracis, Bacterial Proteins, Bacterial Toxins, Carbon Dioxide, Female, Glucose, Virulence, Trans-Activators, Gene Expression Regulation, Bacterial, Virulence Factors, Mice

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