Characterization of a DNA-damage-recognition protein from F9 teratocarcinoma cells, which is inducible by retinoic acid and cyclic AMP.
A nuclear protein that recognizes u.v.-damaged DNA was detected in extracts from murine F9 embryonic stem cells using a DNA-binding assay. The nuclear-protein-binding activity was increased in cells after treatment with retinoic acid/dibutyryl cyclic AMP (dbcAMP), with optimum induction at 6 days. In vitro treatment of nuclear extracts with agents that affect protein conformation (such as urea, Nonidet P40 and Ca2+) slightly modulated the damage-recognition activity. Furthermore, treatment of nuclear extracts with phosphatase dramatically inhibited the binding activity. In addition, damaged-DNA recognition of the nuclear extracts was effectively inhibited by damaged double- and single-stranded DNA. The expression of the nuclear protein with similar characteristics was abundant in HeLa cells and was increased in drug- or u.v.-resistant cells. The findings suggest that the recognition of a u.v.-DNA adduct is modulated, at least in part, by an activity that is induced during retinoic acid/dbcAMP-induced differentiation. These results also imply that the identified damage-recognition protein may be important for the sensitivity or resistance of mammalian cells to DNA damage.
Department of Biochemistry, Chang Gung Medical College, Taoyuan, Taiwan, Republic of China.
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Lin-Chao
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics; D003994:Bucladesine / Q000494:pharmacology; D002454:Cell Differentiation; D002467:Cell Nucleus / Q000737:chemistry; D000242:Cyclic AMP / Q000494:pharmacology*; D004247:DNA / Q000378:metabolism*; D004249:DNA Damage; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D051379:Mice; D009687:Nuclear Proteins / Q000032:analysis / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D013724:Teratoma / Q000737:chemistry*; D014162:Transfection; D014212:Tretinoin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D014466:Ultraviolet Rays
D009687:Nuclear Proteins; D014212:Tretinoin; D003994:Bucladesine; D004247:DNA; D000242:Cyclic AMP