Abstract
Cyclic AMP (cAMP) receptor protein (Crp) is an important transcriptional regulator of Yersinia pestis. Expression of crp increases during pneumonic plague as the pathogen depletes glucose and forms large biofilms within lungs. To better understand control of Y. pestis Crp, we determined a 1.8-Å crystal structure of the protein-cAMP complex. We found that compared to Escherichia coli Crp, C helix amino acid substitutions in Y. pestis Crp did not impact the cAMP dependency of Crp to bind DNA promoters. To investigate Y. pestis Crp-regulated genes during plague pneumonia, we performed RNA sequencing on both wild-type and Δcrp mutant bacteria growing in planktonic and biofilm states in minimal media with glucose or glycerol. Y. pestis Crp was found to dramatically alter expression of hundreds of genes in a manner dependent upon carbon source and growth state. Gel shift assays confirmed direct regulation of the malT and ptsG promoters, and Crp was then linked to Y. pestis growth on maltose as a sole carbon source. Iron regulation genes ybtA and fyuA were found to be indirectly regulated by Crp. A new connection between carbon source and quorum sensing was revealed as Crp was found to regulate production of acyl-homoserine lactones (AHLs) through direct and indirect regulation of genes for AHL synthetases and receptors. AHLs were subsequently identified in the lungs of Y. pestisinfected mice when crp expression was highest in Y. pestis biofilms. Thus, in addition to the well-studied pla gene, other Crp-regulated genes likely have important functions during plague infection. IMPORTANCE Bacterial pathogens have evolved extensive signaling pathways to translate environmental signals into changes in gene expression. While Crp has long been appreciated for its role in regulating metabolism of carbon sources in many bacterial species, transcriptional profiling has revealed that this protein regulates many other aspects of bacterial physiology. The plague pathogen Y. pestis requires this global regulator to survive in blood, skin, and lungs. During disease progression, this organism adapts to changes within these niches. In addition to regulating genes for metabolism of nonglucose sugars, we found that Crp regulates genes for virulence, metal acquisition, and quorum sensing by direct or indirect mechanisms. Thus, this single transcriptional regulator, which responds to changes in available carbon sources, can regulate multiple critical behaviors for causing disease.
Original language | English (US) |
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Article number | e02613-19 |
Journal | mBio |
Volume | 10 |
Issue number | 6 |
DOIs | |
State | Published - Nov 1 2019 |
Funding
We thank Ludmilla Shuvalova and Olga Kiryukhina for expression, purification, crystallization, and data collection of the Crp protein. Use of the Tecan Safire II plate reader and access to structure determination software and LS-CAT were provided by the Northwestern University Structural Biology Facility supported by NCICSG P30-CA60553. Chimera is developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco (supported by NIGMS P41-GM103311). This project was funded in whole or in part with federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services, under contract number HHSN272201700060C and grant number R21 AI111018 (to K.J.F.S.). Use of the Tecan Safire II plate reader and access to structure determination software and LS-CAT were provided by the Northwestern University Structural Biology Facility supported by NCICSG P30-CA60553. Chimera is developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco (supported by NIGMS P41-GM103311). This project was funded in whole or in part with federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services, under contract number HHSN272201700060C and grant number R21 AI111018 (to K.J.F.S.).
Keywords
- Biofilms
- Catabolite repression
- Crp
- Cyclic AMP
- Plague
- Quorum sensing
- RNA-seq
- Regulation of gene expression
- Structure
- Threedimensional structure
- Yersinia pestis
ASJC Scopus subject areas
- Microbiology
- Virology
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1.8 Angstrom Resolution Crystal Structure of cAMP-Regulatory Protein from Yersinia pestis in Complex with cAMP
Ritzert, J. T. (Contributor), Minasov, G. (Contributor), Embry, R. (Contributor), Schipma, M. J. (Contributor) & Satchell, K. J. F. (Contributor), Protein Data Bank (PDB), Jun 27 2018
DOI: 10.2210/pdb6DT4/pdb, https://www.wwpdb.org/pdb?id=pdb_00006dt4
Dataset