Subject(s)
DNA/metabolism , Ethylamines/analysis , Genes, fos , Promoter Regions, Genetic , Pyrimidine Nucleotides/pharmacology , Transcription, Genetic/drug effects , Alkylation , Animals , Base Sequence , Binding Sites , Cell Line , Chloramphenicol O-Acetyltransferase/genetics , DNA Footprinting , Mice , Molecular Sequence Data , Pyrimidine Nucleotides/chemistry , TransfectionABSTRACT
Using gel-retardation assay we have investigated binding of nuclear proteins to the mouse c-fos promoter region 30 b.p. long (nucleotides (-464) - (-435) from TATA-box), localized upstream of PDGF-dependent induction element. It was found that some factors from nuclear extracts of various human and murine cells bind to this promoter region. After gel-retardation of DNA- protein complexes from nuclear extracts of quiescent and stimulated with 20% fetal calf serum cells we observed only one retarded band, while after gel-retardation of DNA-protein complexes from proliferating pseudonormal and tumorigenic cells we observed the appearance of additional retarded band with higher mobility in PAAG. We also have determined the molecular weights of factors interacting with investigated c-fos promoter region by affinity modification method. The molecular weights of both factors are 59 kDa. The equality of molecular weights of investigated factors suggests that these factors might be different forms of one nuclear protein.