High-throughput Sequencing Reveals Suppressors of Vibrio cholerae rpoE Mutations: One Fewer Porin is Enough
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High-throughput Sequencing Reveals Suppressors of Vibrio cholerae rpoE Mutations: One Fewer Porin is Enough
| Title: | High-throughput Sequencing Reveals Suppressors of Vibrio cholerae rpoE Mutations: One Fewer Porin is Enough |
| Author: |
Davis, Brigid; Waldor, Matthew K
Note: Order does not necessarily reflect citation order of authors. |
| Citation: | Davis, Brigid M., and Matthew K. Waldor. 2009. High-throughput sequencing reveals suppressors of Vibrio cholerae rpoE mutations: One fewer porin is enough. Nucleic Acids Research 37(17): 5757-5767. |
| Full Text & Related Files: |
2761261.pdf (1.731Mb; PDF) 
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| Abstract: | Analyses of suppressor mutations have been extremely valuable in understanding gene function. However, techniques for mapping suppressor mutations are not available for most bacterial species. Here, we used high-throughput sequencing technology to identify spontaneously arising suppressor mutations that enabled disruption of rpoE (which encodes σ\(^E\)) in Vibrio cholerae, the agent of cholera. The alternative sigma factor σ\(^E\), which is activated by envelope stress, promotes expression of factors that help preserve and/or restore cell envelope integrity. In Escherichia coli, rpoE is an essential gene that can only be disrupted in the presence of additional suppressor mutations. Among a panel of independent V. cholerae rpoE mutants, more than 75% contain suppressor mutations that reduce production of OmpU, V. cholerae’s principal outer membrane porin. OmpU appears to be a key determinant of V. cholerae’s requirement for and production of σ\(^E\). Such dependence upon a single factor contrasts markedly with regulation of σ\(^E\) in E. coli, in which numerous factors contribute to its activation and none is dominant. We also identified a suppressor mutation that differs from all previously described suppressors in that it elevates, rather than reduces, σ\(^E\)’s activity. Finally, analyses of a panel of rpoE mutants shed light on the mechanisms by which suppressor mutations may arise in V. cholerae. |
| Published Version: | doi://10.1093/nar/gkp568 |
| Other Sources: | http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2761261/pdf/ |
| Terms of Use: | This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAA |
| Citable link to this page: | http://nrs.harvard.edu/urn-3:HUL.InstRepos:5336036 |
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