Overexpression of CupB5 activates alginate overproduction in Pseudomonas aeruginosa by a novel AlgW-dependent mechanism
In Pseudomonas aeruginosa, alginate overproduction, also known as mucoidy, is negatively regulated by the transmembrane protein MucA, which sequesters the alternative sigma factor AlgU. MucA is degraded via a proteolysis pathway that frees AlgU from sequestration, activating alginate biosynthesis. I...
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| Format: | Article |
| Language: | en_US |
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Wiley Blackwell
2017
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| Online Access: | http://hdl.handle.net/1721.1/106340 https://orcid.org/0000-0002-1719-5399 |
| _version_ | 1826202804683800576 |
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| author | Yin, Yeshi Withers, T. Ryan Wang, Xin Yu, Hongwei D. de Regt, Anna Katherine Baker, Tania Sauer, Robert T. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Yin, Yeshi Withers, T. Ryan Wang, Xin Yu, Hongwei D. de Regt, Anna Katherine Baker, Tania Sauer, Robert T. |
| author_sort | Yin, Yeshi |
| collection | MIT |
| description | In Pseudomonas aeruginosa, alginate overproduction, also known as mucoidy, is negatively regulated by the transmembrane protein MucA, which sequesters the alternative sigma factor AlgU. MucA is degraded via a proteolysis pathway that frees AlgU from sequestration, activating alginate biosynthesis. Initiation of this pathway normally requires two signals: peptide sequences in unassembled outer-membrane proteins (OMPs) activate the AlgW protease, and unassembled lipopolysaccharides bind periplasmic MucB, releasing MucA and facilitating its proteolysis by activated AlgW. To search for novel alginate regulators, we screened a transposon library in the non-mucoid reference strain PAO1, and identified a mutant that confers mucoidy through overexpression of a protein encoded by the chaperone-usher pathway gene cupB5. CupB5-dependent mucoidy occurs through the AlgU pathway and can be reversed by overexpression of MucA or MucB. In the presence of activating OMP peptides, peptides corresponding to a region of CupB5 needed for mucoidy further stimulated AlgW cleavage of MucA in vitro. Moreover, the CupB5 peptide allowed OMP-activated AlgW cleavage of MucA in the presence of the MucB inhibitor. These results support a novel mechanism for conversion to mucoidy in which the proteolytic activity of AlgW and its ability to compete with MucB for MucA is mediated by independent peptide signals. |
| first_indexed | 2024-09-23T12:19:27Z |
| format | Article |
| id | mit-1721.1/106340 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T12:19:27Z |
| publishDate | 2017 |
| publisher | Wiley Blackwell |
| record_format | dspace |
| spelling | mit-1721.1/1063402022-10-01T08:59:07Z Overexpression of CupB5 activates alginate overproduction in Pseudomonas aeruginosa by a novel AlgW-dependent mechanism Yin, Yeshi Withers, T. Ryan Wang, Xin Yu, Hongwei D. de Regt, Anna Katherine Baker, Tania Sauer, Robert T. Massachusetts Institute of Technology. Department of Biology de Regt, Anna Katherine Baker, Tania Sauer, Robert T. In Pseudomonas aeruginosa, alginate overproduction, also known as mucoidy, is negatively regulated by the transmembrane protein MucA, which sequesters the alternative sigma factor AlgU. MucA is degraded via a proteolysis pathway that frees AlgU from sequestration, activating alginate biosynthesis. Initiation of this pathway normally requires two signals: peptide sequences in unassembled outer-membrane proteins (OMPs) activate the AlgW protease, and unassembled lipopolysaccharides bind periplasmic MucB, releasing MucA and facilitating its proteolysis by activated AlgW. To search for novel alginate regulators, we screened a transposon library in the non-mucoid reference strain PAO1, and identified a mutant that confers mucoidy through overexpression of a protein encoded by the chaperone-usher pathway gene cupB5. CupB5-dependent mucoidy occurs through the AlgU pathway and can be reversed by overexpression of MucA or MucB. In the presence of activating OMP peptides, peptides corresponding to a region of CupB5 needed for mucoidy further stimulated AlgW cleavage of MucA in vitro. Moreover, the CupB5 peptide allowed OMP-activated AlgW cleavage of MucA in the presence of the MucB inhibitor. These results support a novel mechanism for conversion to mucoidy in which the proteolytic activity of AlgW and its ability to compete with MucB for MucA is mediated by independent peptide signals. United States. National Aeronautics and Space Administration. West Virginia Space Grant Consortium Canadian Cystic Fibrosis Foundation (Grant CFF-YU11G0) National Institutes of Health (U.S.) (Grant AI-16892) 2017-01-11T16:14:44Z 2017-01-11T16:14:44Z 2014-07 2014-06 Article http://purl.org/eprint/type/JournalArticle 0950-382X 1365-2958 http://hdl.handle.net/1721.1/106340 de Regt, Anna K. et al. “Overexpression of CupB5 Activates Alginate Overproduction in P Seudomonas Aeruginosa by a Novel AlgW-Dependent Mechanism: CupB5 Activates Alginate Production through AlgW.” Molecular Microbiology 93.3 (2014): 415–425. https://orcid.org/0000-0002-1719-5399 en_US http://dx.doi.org/10.1111/mmi.12665 Molecular Microbiology Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Wiley Blackwell PMC |
| spellingShingle | Yin, Yeshi Withers, T. Ryan Wang, Xin Yu, Hongwei D. de Regt, Anna Katherine Baker, Tania Sauer, Robert T. Overexpression of CupB5 activates alginate overproduction in Pseudomonas aeruginosa by a novel AlgW-dependent mechanism |
| title | Overexpression of CupB5 activates alginate overproduction in Pseudomonas aeruginosa by a novel AlgW-dependent mechanism |
| title_full | Overexpression of CupB5 activates alginate overproduction in Pseudomonas aeruginosa by a novel AlgW-dependent mechanism |
| title_fullStr | Overexpression of CupB5 activates alginate overproduction in Pseudomonas aeruginosa by a novel AlgW-dependent mechanism |
| title_full_unstemmed | Overexpression of CupB5 activates alginate overproduction in Pseudomonas aeruginosa by a novel AlgW-dependent mechanism |
| title_short | Overexpression of CupB5 activates alginate overproduction in Pseudomonas aeruginosa by a novel AlgW-dependent mechanism |
| title_sort | overexpression of cupb5 activates alginate overproduction in pseudomonas aeruginosa by a novel algw dependent mechanism |
| url | http://hdl.handle.net/1721.1/106340 https://orcid.org/0000-0002-1719-5399 |
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