Transcription factors NF-YA regulate the induction of human OGG1 following DNA-alkylating agent methylmethane sulfonate (MMS) treatment.
A human 8-oxoguanine-DNA glycosylase (hOGG1) is the main enzyme that repairs 8-oxoG, which is a critical mutagenic lesion. There is a great deal of interest in the up- or down-regulation of OGG1 expression after DNA damage. In this study, we investigated the effect of a DNA-alkylating agent, methylmethane sulfonate (MMS), on hOGG1 expression level and found that MMS treatment resulted in an increase in the functional hOGG1 expression in HCT116 cells. A region between -121 and -61 of the hOGG1 promoter was found to be crucial for this induction by MMS. Site-directed mutations of the two inverted CCAAT motifs substantially abrogated the induction of the hOGG1 promoter as a result of MMS treatment. In addition, the NF-YA protein (binding to the inverted CCAAT box) was induced after exposing cells to MMS. Moreover, gel shift and supershift analyses with the nuclear extracts prepared from HCT116 cells identified NF-YA as the transcription factor interacting with the inverted CCAAT box. Mutations of the inverted CCAAT box either prevented the binding of this factor or abolished its activation as a result of MMS treatment. Finally, this study showed that hOGG1-expressing HCT116 cells exhibited increased hOGG1 repair activity and resistance to MMS. Overall, these results demonstrate that MMS can up-regulate hOGG1 expression through the induction of the transcription factor, NF-YA, and increased transcription level of the hOGG1 gene correlates with an increase in enzyme activity providing functional protection from MMS.
Lee
Mi-Rha
MR
Research Center for Proteineous Materials, Chosun University, 375 Seosuk-dong, Gwangju 501-759, Korea.
Kim
Soo-Hyun
SH
Cho
Hyun-Ju
HJ
Lee
Kun-Yeong
KY
Moon
Ae Ran
AR
Jeong
Hye Gwang
HG
Lee
Jung-Sup
JS
Hyun
Jin-Won
JW
Chung
Myung-Hee
MH
You
Ho Jin
HJ
D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000477:Alkylating Agents / Q000494:pharmacology*; D020816:Amino Acid Motifs; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology; D015153:Blotting, Western; D023081:CCAAT-Binding Factor / Q000378:metabolism / Q000502:physiology*; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival; D004247:DNA / Q000737:chemistry*; D045647:DNA Glycosylases / Q000378:metabolism*; D004260:DNA Repair; D004305:Dose-Response Relationship, Drug; D015972:Gene Expression Regulation, Neoplastic; D006147:Guanine / Q000031:analogs & derivatives / Q000737:chemistry; D006151:Guanosine / Q000031:analogs & derivatives* / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008156:Luciferases / Q000378:metabolism; D008741:Methyl Methanesulfonate / Q000494:pharmacology*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism / Q000502:physiology*; D014162:Transfection; D015854:Up-Regulation
D000276:Adjuvants, Immunologic; D000477:Alkylating Agents; D018906:Antineoplastic Agents, Alkylating; D023081:CCAAT-Binding Factor; C492061:NFYA protein, human; D014157:Transcription Factors; D006151:Guanosine; C046215:8-hydroxyguanosine; D006147:Guanine; C008450:7-methylguanine; D004247:DNA; D008741:Methyl Methanesulfonate; D006861:Hydrogen Peroxide; D008156:Luciferases; D045647:DNA Glycosylases; C479058:oxoguanine glycosylase 1, human