The Bifunctional Cell Wall Hydrolase CwlT Is Needed for Conjugation of the Integrative and Conjugative Element ICEBs1 in Bacillus subtilis and B. anthracis
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Author(s) •
DeWitt, Tyler
Grossman, Alan Davis
Date Issued
February 2014
Journal
Journal of Bacteriology
Publisher
American Society for Microbiology
Citation
DeWitt, T., and A. D. Grossman. “The Bifunctional Cell Wall Hydrolase CwlT Is Needed for Conjugation of the Integrative and Conjugative Element ICEBs1 in Bacillus Subtilis and B. Anthracis.” Journal of Bacteriology 196, no. 8 (February 14, 2014): 1588–1596.
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Author's final manuscript
Abstract
The mobile genetic element ICEBs1 is an integrative and conjugative element (ICE) found in Bacillus subtilis. One of the ICEBs1 genes, cwlT, encodes a cell wall hydrolase with two catalytic domains, a muramidase and a peptidase. We found that cwlT is required for ICEBs1 conjugation. We examined the role of each of the two catalytic domains and found that the muramidase is essential, whereas the peptidase is partially dispensable for transfer of ICEBs1. We also found that the putative signal peptide in CwlT is required for CwlT to function in conjugation, consistent with the notion that CwlT is normally secreted from the cytoplasm. We found that alteration of the putative lipid attachment site on CwlT had no effect on its role in conjugation, indicating that if CwlT is a lipoprotein, the lipid attachment is not required for conjugation. Finally, we found conditions supporting efficient transfer of ICEBs1 into and out of Bacillus anthracis and that cwlT was needed for ICEBs1 to function in B. anthracis. The mature cell wall of B. anthracis is resistant to digestion by CwlT, indicating that CwlT might act during cell wall synthesis, before modifications of the peptidoglycan are complete.
MIT Department
Massachusetts Institute of Technology. Department of Biology
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DOI of Published Version
https://doi.org/10.1128/jb.00012-14