Chemoresistance is a major problem in cancer therapy, but it is largely unclear how cancer cells become resistant. Deficiency in the mismatch-repair (MMR) proteins allows cells to resist toxic chemotherapeutic agents such as DNA methylators. However, the distinct MMR function that sensitizes cells to DNA methylation is not clearly defined given the multiple MMR functions. MMR proteins function to repair base mismatches after DNA replication, inhibit recombination between non-identical DNA sequences, as well as activate both checkpoint and apoptotic responses following DNA damage. Separation-of- function mutants, suggest that resistance to DNA methylation is dependent on a disrupted checkpoint. Recently, we established that both MMR proteins of the MutL1 complex (MLH1/PMS2) and BACH1/FANCJ (BRCA1-associated C- terminal helicase/Fanconi Anemia complementation group J) are required for checkpoint signaling. Specifically, we identified that BACH1 binds directly to MLH1 and that a mutant version of BACH1 ablated for MLH1 binding failed to elicit an interstrand crosslink (ICL)-induced checkpoint response. Since ICLs activate the intra S-phase checkpoint, and both MLH1 and BACH1 have been shown independently to function in the intra S-phase checkpoint, this checkpoint likely requires the formation of a BACH1/MutL1 complex. We will test this possibility directly. In addition, we will determine whether BACH1 also participates in the DNA-methylation-induced G2/M accumulation checkpoint similar to MutL1. Consistent with a role for BACH1 in the DNA methylation-induced response, our lab has shown that similar to MutL1 deficient cells, BACH1 deficient cells are resistant to DNA methylation. In contrast, BRCA1 deficient cells are sensitive to DNA methylation, suggesting that BACH1 uniquely functions in the DNA methylati1n response. We propose to dissect the role of a BACH1/MutL1 complex in both checkpoint and repair functions. We will determine whether the formation of an intact complex is required to restore chemosensitivity to resistant null BACH1 or MutL1 cells. Defining the function of the BACH1/MutL1 complex ideally will provide insight towards restoring chemosensitivity to cancer cells. Along these lines, we will test whether manipulation of the recombination function of the BACH1/BRCA1 complex will uniquely sensitizes MMR deficient cells to chemotherapies.
