The impact of YabG mutations on C. difficile spore germination and processing of spore substrates.

Autor: Osborne MS; Department of Biology, Texas A&M University, College Station, TX 77845, USA., Brehm JN; Department of Biology, Texas A&M University, College Station, TX 77845, USA., Olivença C; Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal., Cochran AM; Department of Biology, Texas A&M University, College Station, TX 77845, USA., Serrano M; Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal., Henriques AO; Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal., Sorg JA; Department of Biology, Texas A&M University, College Station, TX 77845, USA.
Jazyk: angličtina
Zdroj: BioRxiv : the preprint server for biology [bioRxiv] 2024 Jun 10. Date of Electronic Publication: 2024 Jun 10.
DOI: 10.1101/2024.06.10.598338
Abstrakt: YabG is a sporulation-specific protease that is conserved among sporulating bacteria. C. difficile YabG processes cortex destined proteins preproSleC into proSleC and CspBA to CspB and CspA. YabG also affects synthesis of spore coat/exosporium proteins CotA and CdeM. In prior work that identified CspA as the co-germinant receptor, mutations in yabG were found which altered the co-germinants required to initiate spore germination. To understand how these mutations in the yabG locus contribute to C. difficile spore germination, we introduced these mutations into an isogenic background. Spores derived from C. difficile yabG C207A (catalytically inactive), C. difficile yabG A46D , C. difficile yabG G37E , and C. difficile yabG P153L strains germinated in response to TA alone. Recombinantly expressed and purified preproSleC incubated with E. coli lysate expressing wild type YabG resulted in the removal of the pre sequence from preproSleC. Interestingly, only YabG A46D showed any activity towards purified preproSleC. Mutation of the YabG processing site in preproSleC (R119A) led to YabG shifting its processing to R115 or R112. Finally, changes in yabG expression under the mutant promoters were analyzed using a SNAP-tag and revealed expression differences at early and late stages of sporulation. Overall, our results support and expand upon the hypothesis that YabG is important for germination and spore assembly and, upon mutation of the processing site, can shift where it cleaves substrates.
Competing Interests: Conflict of interest The authors declare no conflict of interest.
Databáze: MEDLINE
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