DNA interstrand crosslinks (ICLs) are among the most toxic types of damage to a cell. For this reason, many ICL-inducing agents are effective therapeutic agents. For example, cisplatin and nitrogen mustards are used for treating cancer and psoralen plus UVA (PUVA) is useful for treating psoriasis. However, repair mechanisms for ICLs in the human genome are not clearly defined. Previously, we have shown that MSH2, the common subunit of the human MutSα and MutSβ mismatch recognition complexes, plays a role in the error-free repair of psoralen ICLs. We hypothesized that MLH1, the common subunit of human MutL complexes, is also involved in the cellular response to psoralen ICLs. Surprisingly, we instead found that MLH1-deficient human cells are more resistant to psoralen ICLs, in contrast to the sensitivity to these lesions displayed by MSH2-deficient cells. Apoptosis was not as efficiently induced by psoralen ICLs in MLH1-deficient cells as in MLH1-proficient cells as determined by caspase-3/7 activity and binding of annexin V. Strikingly, CHK2 phosphorylation was undetectable in MLH1-deficient cells, and phosphorylation of CHK1 was reduced after PUVA treatment, indicating that MLH1 is involved in signaling psoralen ICL-induced checkpoint activation. Psoralen ICLs can result in mutations near the crosslinked sites; however, MLH1 function was not required for the mutagenic repair of these lesions, and so its signaling function appears to have a role in maintaining genomic stability following exposure to ICL-induced DNA damage. Distinguishing the genetic status of MMR-deficient tumors as MSH2-deficient or MLH1-deficient is thus potentially important in predicting the efficacy of treatment with psoralen and perhaps with other ICL-inducing agents.
Crosslinks, linking the complementary stands of the DNA double helix, can lead to cell death, because they are so effective at interfering with normal genomic transactions such as DNA replication. This property of crosslinking agents has long been utilized in cancer therapy. The purpose of our research is to understand the function of DNA repair proteins in cellular responses to DNA interstrand crosslinking agents. MSH2 is a central protein in the recognition of DNA mismatches, and we previously found that it plays an important role in protecting cells against the toxicity of crosslinks. The MLH1 protein functions in DNA mismatch repair in a later step, and we hypothesized that MLH1 may also be involved in repair of crosslinks. We were surprised to find that MLH1 function is important for DNA crosslink-induced signaling, rather than DNA repair. MLH1-deficient cells are more resistant to crosslinks and have defective signaling to processes that signal cell death. This work may have clinical consequences, as mutations in MSH2 and MLH1 are common in tumors. MSH2-deficient cells may be more vulnerable to DNA crosslink-inducing agents than normal, while MLH1-deficient cells have a greater potential to survive crosslinking treatment, which could instead potentiate further tumor initiation.