The Fanconi Anemia Core Complex Forms Four Complexes of Different Sizes in Different Subcellular Compartments

摘要: Fanconi anemia (FA) is an autosomal recessive disease marked by congenital defects, bone marrow failure, and cancer susceptibility. FA cells exhibit a characteristic hypersensitivity to DNA crosslinking agents such as mitomycin C. The molecular mechanism for the remains elusive, but at least 6 proteins are known be part of what termed core complex. We used affinity pulldown FLAG-FANCA pull down complex from whole-cell extracts. Mass spectroscopy detected previously reported FA-binding proteins, including FANCA, FANCC, FANCG, cdc2, GRP94, thus validating approach. further describe method purification in effort find novel components biochemical activity define function By using conventional chromatographic fractionation subcellular preparations, we report: (i) exists cytoplasmic form 500-600 kDa; (ii) larger, 750-kDa seen only mitosis; (iii) nuclear achieves size 2 megaDaltons; (iv) distinct 1-megaDalton bound chromatin that contains phosphorylated FANCA after undergoing damage. continuing our analysis mass characterize binding proteins. These data will help role normal cell physiology well development state.