A patient-derived mutant form of the Fanconi anemia protein, FANCA, is defective in nuclear accumulation

Fanconi anemia (FA) is an autosomal recessive cancer susceptibility syndrome with at least eight complementation groups (A-H). Three FA genes, corresponding to complementation groups A, C, and G, have been cloned, but the function of the encoded FA proteins remains unknown. We recently demonstrated that the FANCA and FANCC proteins bind and form a nuclear complex, In the current study, we identified a homozygous mutation in the FANCA gene (3329A>C) in an Egyptian FA patient from a consanguineous family. This mutant FANCA allele is predicted to encode a mutant FANCA protein, FANCA(H1110P), in which histidine 1110 is changed to proline, Initially, me characterized the FANCA(H1110P) protein, expressed in an Epstein Parr virus (EBV)-immortalized lymphoblast Line derived from the patient. Unlike wild-type FANCA protein expressed in normal lymphoblasts, FANCA(H1110P) was not phosphorylated and failed to bind to FANCC. To test directly the effect of this mutation on FANCA function, we used retroviral-mediated transduction to express either wildtype FANCA or FANCA(H1110P) protein in the FA-A fibroblast line, GM6914. Unlike mild-type FANCA, the mutant protein failed to complement the mitomycin C sensitivity of these cells, In addition, the FANCA(H1110P) protein mas defective in nuclear accumulation in the transduced cells. The characteristics of this mutant protein underscore the importance of FANCA phosphorylation, FANCA/FANCC binding, and nuclear accumulation in the function of the FA pathway. (C) 1999 International Society for Experimental Hematology. Published by Elsevier Science Inc.