A Drosophila Model for Clostridium difficile Toxin CDT Reveals Interactions with Multiple Effector Pathways
Summary: Clostridium difficile infections (CDIs) cause severe and occasionally life-threatening diarrhea. Hyper-virulent strains produce CDT, a toxin that ADP-ribosylates actin monomers and inhibits actin polymerization. We created transgenic Drosophila lines expressing the catalytic subunit CDTa to...
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| Format: | Article |
| Language: | English |
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Elsevier
2020-02-01
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| Series: | iScience |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004220300481 |
| _version_ | 1819051496074903552 |
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| author | Ruth Schwartz Annabel Guichard Nathalie C. Franc Sitara Roy Ethan Bier |
| author_facet | Ruth Schwartz Annabel Guichard Nathalie C. Franc Sitara Roy Ethan Bier |
| author_sort | Ruth Schwartz |
| collection | DOAJ |
| description | Summary: Clostridium difficile infections (CDIs) cause severe and occasionally life-threatening diarrhea. Hyper-virulent strains produce CDT, a toxin that ADP-ribosylates actin monomers and inhibits actin polymerization. We created transgenic Drosophila lines expressing the catalytic subunit CDTa to investigate its interaction with host signaling pathways in vivo. When expressed in the midgut, CDTa reduces body weight and fecal output and compromises survival, suggesting severe impairment of digestive functions. At the cellular level, CDTa induces F-actin network collapse, elimination of the intestinal brush border, and disruption of intercellular junctions. We confirm toxin-dependent re-distribution of Rab11 to enterocytes' apical surface and observe suppression of CDTa phenotypes by a Dominant-Negative form of Rab11 or RNAi of the dedicated Rab11GEF Crag (DENND4). We also report that Calmodulin (Cam) is required to mediate CDTa activity. In parallel, chemical inhibition of the Cam/Calcineurin pathway by Cyclosporin A or FK506 also reduces CDTa phenotypes, potentially opening new avenues for treating CDIs. : Molecular Genetics; Model Organism Subject Areas: Molecular Genetics, Model Organism |
| first_indexed | 2024-12-21T12:04:52Z |
| format | Article |
| id | doaj.art-6e070d472ea2492aa8c0e6eae8f8f453 |
| institution | Directory Open Access Journal |
| issn | 2589-0042 |
| language | English |
| last_indexed | 2024-12-21T12:04:52Z |
| publishDate | 2020-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj.art-6e070d472ea2492aa8c0e6eae8f8f4532022-12-21T19:04:43ZengElsevieriScience2589-00422020-02-01232A Drosophila Model for Clostridium difficile Toxin CDT Reveals Interactions with Multiple Effector PathwaysRuth Schwartz0Annabel Guichard1Nathalie C. Franc2Sitara Roy3Ethan Bier4Section of Cell and Developmental Biology, University of California, San Diego, La Jolla, CA 92093-0335, USASection of Cell and Developmental Biology, University of California, San Diego, La Jolla, CA 92093-0335, USA; Tata Institute for Genetics and Society-UCSD, La Jolla, CA 92093-0335, USAFranc Consulting, San Diego, CA 92117-3314, USA; The Scripps Research Institute, La Jolla, CA 92037, USASection of Cell and Developmental Biology, University of California, San Diego, La Jolla, CA 92093-0335, USASection of Cell and Developmental Biology, University of California, San Diego, La Jolla, CA 92093-0335, USA; Tata Institute for Genetics and Society-UCSD, La Jolla, CA 92093-0335, USA; Corresponding authorSummary: Clostridium difficile infections (CDIs) cause severe and occasionally life-threatening diarrhea. Hyper-virulent strains produce CDT, a toxin that ADP-ribosylates actin monomers and inhibits actin polymerization. We created transgenic Drosophila lines expressing the catalytic subunit CDTa to investigate its interaction with host signaling pathways in vivo. When expressed in the midgut, CDTa reduces body weight and fecal output and compromises survival, suggesting severe impairment of digestive functions. At the cellular level, CDTa induces F-actin network collapse, elimination of the intestinal brush border, and disruption of intercellular junctions. We confirm toxin-dependent re-distribution of Rab11 to enterocytes' apical surface and observe suppression of CDTa phenotypes by a Dominant-Negative form of Rab11 or RNAi of the dedicated Rab11GEF Crag (DENND4). We also report that Calmodulin (Cam) is required to mediate CDTa activity. In parallel, chemical inhibition of the Cam/Calcineurin pathway by Cyclosporin A or FK506 also reduces CDTa phenotypes, potentially opening new avenues for treating CDIs. : Molecular Genetics; Model Organism Subject Areas: Molecular Genetics, Model Organismhttp://www.sciencedirect.com/science/article/pii/S2589004220300481 |
| spellingShingle | Ruth Schwartz Annabel Guichard Nathalie C. Franc Sitara Roy Ethan Bier A Drosophila Model for Clostridium difficile Toxin CDT Reveals Interactions with Multiple Effector Pathways iScience |
| title | A Drosophila Model for Clostridium difficile Toxin CDT Reveals Interactions with Multiple Effector Pathways |
| title_full | A Drosophila Model for Clostridium difficile Toxin CDT Reveals Interactions with Multiple Effector Pathways |
| title_fullStr | A Drosophila Model for Clostridium difficile Toxin CDT Reveals Interactions with Multiple Effector Pathways |
| title_full_unstemmed | A Drosophila Model for Clostridium difficile Toxin CDT Reveals Interactions with Multiple Effector Pathways |
| title_short | A Drosophila Model for Clostridium difficile Toxin CDT Reveals Interactions with Multiple Effector Pathways |
| title_sort | drosophila model for clostridium difficile toxin cdt reveals interactions with multiple effector pathways |
| url | http://www.sciencedirect.com/science/article/pii/S2589004220300481 |
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