Identification of ClpP Dual Isoform Disruption as an Antisporulation Strategy for Clostridioides difficile
| العنوان: | Identification of ClpP Dual Isoform Disruption as an Antisporulation Strategy for Clostridioides difficile |
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| المؤلفون: | Jimmy D. Ballard, Adam S Duerfeldt, Tyler M. Shadid, Catherine E Bishop, Megan L. Kempher, Nathan P. Lavey |
| المصدر: | J Bacteriol |
| بيانات النشر: | American Society for Microbiology, 2022. |
| سنة النشر: | 2022 |
| مصطلحات موضوعية: | medicine.medical_treatment, Mutant, Druggability, Virulence, Biology, Microbiology, Bortezomib, Bacterial Proteins, Genome editing, medicine, Humans, Protein Isoforms, Coding region, Molecular Biology, Gene Editing, Spores, Bacterial, Protease, Clostridioides difficile, Gene Expression Regulation, Bacterial, Phenotype, Stop codon, Mutation, Clostridium Infections, Research Article |
| الوصف: | The Gram-positive bacterium Clostridioides difficile is a primary cause of hospital-acquired diarrhea, threatening both immunocompromised and healthy individuals. An important aspect of defining mechanisms that drive C. difficile persistence and virulence relies on developing a more complete understanding of sporulation. C. difficile sporulation is the single determinant of transmission and complicates treatment and prevention due to the chemical and physical resilience of spores. By extension, the identification of druggable targets that significantly attenuate sporulation would have a significant impact on thwarting C. difficile infection. By use of a new CRISPR-Cas9 nickase genome editing methodology, stop codons were inserted early in the coding sequence for clpP1 and clpP2 to generate C. difficile mutants that no longer produced the corresponding isoforms of caseinolytic protease P (ClpP). The data show that genetic ablation of ClpP isoforms leads to altered sporulation phenotypes with the clpP1/clpP2 double mutant exhibiting asporogenic behavior. A small screen of known ClpP inhibitors in a fluorescence-based biochemical assay identified bortezomib as an inhibitor of C. difficile ClpP that produces dose-dependent inhibition of purified ClpP. Incubation of C. difficile cultures in the presence of bortezomib reveals antisporulation effects approaching that observed in the clpP1/clpP2 double mutant. This work identifies ClpP as a key contributor to C. difficile sporulation and provides compelling support for the pursuit of small-molecule ClpP inhibitors as C. difficile antisporulating agents. IMPORTANCE Due to diverse roles of ClpP and the reliance of pathogens upon this system for infection, it has emerged as a target for antimicrobial development. Biology regulated by ClpP is organism dependent and has not been defined in Clostridioides difficile. This work identifies ClpP as a key contributor to C. difficile sporulation and provides compelling support for the pursuit of small-molecule ClpP inhibitors as antisporulating agents. The identification of new approaches and/or drug targets that reduce C. difficile sporulation would be transformative and are expected to find high utility in prophylaxis, transmission attenuation, and relapse prevention. Discovery of the ClpP system as a major driver to sporulation also provides a new avenue of inquiry for advancing the understanding of sporulation. |
| تدمد: | 1098-5530 0021-9193 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0d89e26f317c6077b2567f28c48fbb15 https://doi.org/10.1128/jb.00411-21 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....0d89e26f317c6077b2567f28c48fbb15 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10985530 00219193 |
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