Journal article
Identification and characterization of a gene cluster required for proper rod shape, cell division, and pathogenesis in Clostridium difficile
Journal of bacteriology, Vol.196(12), pp.2290-2300
06/2014
DOI: 10.1128/JB.00038-14
PMCID: PMC4054185
PMID: 24727226
Abstract
Little is known about cell division in Clostridium difficile, a strict anaerobe that causes serious diarrheal diseases in people whose normal intestinal microbiome has been perturbed by treatment with broad-spectrum antibiotics. Here we identify and characterize a gene cluster encoding three cell division proteins found only in C. difficile and a small number of closely related bacteria. These proteins were named MldA, MldB, and MldC, for midcell localizing division proteins. MldA is predicted to be a membrane protein with coiled-coil domains and a peptidoglycan-binding SPOR domain. MldB and MldC are predicted to be cytoplasmic proteins; MldB has two predicted coiled-coil domains, but MldC lacks obvious conserved domains or sequence motifs. Mutants of mldA or mldB had morphological defects, including loss of rod shape (a curved cell phenotype) and inefficient separation of daughter cells (a chaining phenotype). Fusions of cyan fluorescent protein (CFP) to MldA, MldB, and MldC revealed that all three proteins localize sharply to the division site. This application of CFP was possible because we discovered that O2-dependent fluorescent proteins produced anaerobically can acquire fluorescence after cells are fixed with cross-linkers to preserve native patterns of protein localization. Mutants lacking the Mld proteins are severely attenuated for pathogenesis in a hamster model of C. difficile infection. Because all three Mld proteins are essentially unique to C. difficile, they might be exploited as targets for antibiotics that combat C. difficile without disrupting the intestinal microbiome.
Details
- Title: Subtitle
- Identification and characterization of a gene cluster required for proper rod shape, cell division, and pathogenesis in Clostridium difficile
- Creators
- Eric M Ransom - Department of Microbiology, University of Iowa, Iowa City, Iowa, USAKyle B Williams - Department of Microbiology, University of Iowa, Iowa City, Iowa, USADavid S Weiss - Department of Microbiology, University of Iowa, Iowa City, Iowa, USA david-weiss@uiowa.edu craig-ellermeier@uiowa.eduCraig D Ellermeier - Department of Microbiology, University of Iowa, Iowa City, Iowa, USA david-weiss@uiowa.edu craig-ellermeier@uiowa.edu
- Resource Type
- Journal article
- Publication Details
- Journal of bacteriology, Vol.196(12), pp.2290-2300
- Publisher
- United States
- DOI
- 10.1128/JB.00038-14
- PMID
- 24727226
- PMCID
- PMC4054185
- ISSN
- 0021-9193
- eISSN
- 1098-5530
- Grant note
- T32 AI007511 / NIAID NIH HHS R01 AI087834 / NIAID NIH HHS P51 OD011132 / NIH HHS T32 AI07511 / NIAID NIH HHS R01 GM083975 / NIGMS NIH HHS GM-083975 / NIGMS NIH HHS
- Language
- English
- Date published
- 06/2014
- Academic Unit
- Microbiology and Immunology
- Record Identifier
- 9984001220502771
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