FANCG binding prolongs the cellular half-life of the FANCA protein.
(A) EUFA316(FA-G) cells or EUFA316 cells stably transfected and corrected with FANCG cDNA were metabolically labeled with35S-methionine for 30 minutes, washed, and chased in cold (unlabeled) medium for the indicated time periods. The cells were lysed, and labeled proteins were immunoprecipitated with an affinity-purified anti-FANCA antiserum. The indicated bands (endogenous FANCA protein) were scanned, and densitometric values were plotted to quantify protein stability. Wild type lymphoblasts express comparable levels of endogenous FANCA protein (data not shown). (B) HSC72(FA-A) cells or HSC72 cells stably transfected and corrected with the FANCA cDNA were analyzed. Labeled proteins were immunoprecipitated with an affinity-purified anti-FANCG antiserum. (C) PD-4(FA-C) cells or PD4 cells stably transfected and corrected with the FANCC cDNA were metabolically labeled, and FANCA and FANCG proteins were analyzed.
