Characterization of a protein recognizing minor groove binders-damaged DNA.
By using electromobility shift assay (EMSA), we have identified a protein able to recognize the DNA only if it was previously reacted with minor groove binders. This protein binds with very high affinity AT containing DNA treated with minor groove binders such as distamycin A, Hoechst 33258 and 33342, CC-1065 and ethidium bromide minor groove intercalator, but not with major groove binders such as quinacrine mustard, cisplatin or melphalan, or with topoisomerase I inhibitor camptothecin or topoisomerase II inhibitor doxorubicin. This protein was found to be present in different extracts of human, murine and hamster cells, with the human protein which appears to have a molecular weight slightly lower than that of the other species. This protein was found to be expressed both in cancer and normal tissues. By using molecular ultrafiltration techniques as well as southwestern analysis it was estimated that the apparent molecular weight is close to 100 kDa. We can exclude an identity between this protein and other proteins, with a similar molecular weight previously reported to be involved in DNA damage recognition/repair, such as topoisomerase I, mismatch repair activities such as the prokaryotic MutS protein and its human homologue hMSH2 or proteins of the nucleotide excision repair system such as ERCC1, -2, -3 and -4.
Colella
G
G
Molecular Pharmacology Unit, LCP, Department of Oncology, Istituto di Richerche Farmacologiche Mario Negri, Milan, Italy.
Bonfanti
M
M
D'Incalci
M
M
Broggini
M
M
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001665:Binding Sites; D006690:Bisbenzimidazole / Q000494:pharmacology; D002460:Cell Line; D006224:Cricetinae; D004247:DNA / Q000187:drug effects / Q000378:metabolism; D004249:DNA Damage; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004214:Distamycins / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007364:Intercalating Agents / Q000494:pharmacology; D019169:Jurkat Cells; D008558:Melphalan / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D009588:Nitrogen Mustard Compounds / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D004268:DNA-Binding Proteins; D004214:Distamycins; D007364:Intercalating Agents; D009588:Nitrogen Mustard Compounds; C066900:tallimustine; C030000:stallimycin; D004247:DNA; D006690:Bisbenzimidazole; D008558:Melphalan