MMR/c-Abl-dependent activation of ING2/p73alpha signaling regulates the cell death response to N-methyl-N'-nitro-N-nitrosoguanidine.
Agents inducing O(6)-methylguanine (O(6)MeG) in DNA such as N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) are cytotoxic and a deficiency in mismatch repair (MMR) results in lack of sensitivity to this genotoxin (termed alkylation tolerance). Here, we show that ING2, a member of the inhibitor of growth family, is required for cell death induced by MNNG. We further observe that MNNG treatment increases cellular protein levels of ING2 that is dependent on intact MMR function and that MNNG-induced ING2 localizes and associates with p73alpha in the nucleus. Suppression of ING2 by short hairpin RNA (shRNA) in MMR-proficient colorectal cancer cells decreased its sensitivity to MNNG and, in addition, abrogated MNNG-induced stabilization and acetylation of p73alpha. Interestingly, suppression of p73alpha had a greater impact on MNNG-induced cell death than ING2 leading us to conclude that ING2 regulates the cell death response, in part, through p73alpha. Inhibition of c-Abl by STI571 or suppression of c-Abl expression by shRNA blocked ING2 induction and p73alpha acetylation induced by this alkylator. Similarly, suppression of MMR (MLH1) by shRNA abrogated ING2 induction/p73alpha acetylation. Taken together, these results demonstrate that MLH1/c-Abl-dependent activation of ING2>p73alpha signaling regulates cell death triggered by MNNG and further suggests that dysregulation of this event may, in part, be responsible for alkylation tolerance observed in MMR compromised cells.
Sun
Guoming
G
Department of Microbiology and Molecular Genetics, School of Medicine, University of Pittsburgh, E1205 Biomedical Science Tower, Pittsburgh, PA 15261, USA.
Jin
Shunqian
S
Baskaran
R
R
D048868:Adaptor Proteins, Signal Transducing / Q000187:drug effects / Q000378:metabolism; D000477:Alkylating Agents / Q000633:toxicity*; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis; D001549:Benzamides; D002273:Carcinogens / Q000633:toxicity*; D053843:DNA Mismatch Repair; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006367:HeLa Cells; D018398:Homeodomain Proteins / Q000819:agonists / Q000378:metabolism*; D006801:Humans; D000068877:Imatinib Mesylate; D008769:Methylnitronitrosoguanidine / Q000633:toxicity*; D000070957:MutL Protein Homolog 1; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000378:metabolism*; D009841:Oligonucleotides / Q000378:metabolism; D010879:Piperazines / Q000494:pharmacology; D016315:Proto-Oncogene Proteins c-abl / Q000187:drug effects / Q000378:metabolism*; D011743:Pyrimidines / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000819:agonists / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D014162:Transfection; D000072160:Tumor Protein p73; D016159:Tumor Suppressor Protein p53 / Q000187:drug effects / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*
D048868:Adaptor Proteins, Signal Transducing; D000477:Alkylating Agents; D000970:Antineoplastic Agents; D001549:Benzamides; D002273:Carcinogens; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; C444654:ING2 protein, human; C501486:MLH1 protein, human; D009687:Nuclear Proteins; D009841:Oligonucleotides; D010879:Piperazines; D011743:Pyrimidines; D034741:RNA, Small Interfering; D018160:Receptors, Cytoplasmic and Nuclear; D000072160:Tumor Protein p73; D016159:Tumor Suppressor Protein p53; D025521:Tumor Suppressor Proteins; D008769:Methylnitronitrosoguanidine; D000068877:Imatinib Mesylate; D016315:Proto-Oncogene Proteins c-abl; D000070957:MutL Protein Homolog 1