Amy, CampMorrison, Jade2025-07-152025-07-15https://hdl.handle.net/10166/6774Protein degradation is a tightly regulated mechanism in bacterial cells that ensures that the correct proteins are being degraded appropriately to avoid harmful consequences within the cell. ClpCP is an example of an AAA+ protease that facilitates proteolysis within bacterial cells through a degron tag or with the assistance of an adaptor protein. MdfA is a recently discovered sporulation-specific adaptor protein that interacts with ClpCP within the bacterium, Bacillus subtilis. Based on results from a previous study, it is speculated that MdfA interacts with the ClpC-M domain, corresponding with an AlphaFold Multimer structure that collaborators constructed. This study investigates whether the ClpC M-domain is a site of interaction for MdfA, which was tested by mutating the appropriate codon of residues located at the contact point between the M-domain and MdfA C-terminal domain. A series of bacterial two-hybrid assays has demonstrated that the M-domain is a potential interaction site and that the ClpC M-domain residues are more critical for MdfA than MecA. The results and a parallel study by a Camp lab member, Jen Butler, support the hypothesis that the M-domain is a secondary interaction site for MdfA.en-USMolecular BiologyThe AAA+ ATPase ClpC-M Domain is a Secondary Interaction Site For the Sporulation-Specific Adaptor Protein MdfA in the Bacterium Bacillus SubtilisThesisrestricted