MAINTAINING DNA REPLICATION FORK STABILITY: ROLE OF THE FANCONI ANEMIA PATHWAY

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Fanconi anemia (FA) is a rare disease characterized by congenital anomalies, bone marrow failure, and cancer susceptibility. FA cells are hypersensitive to DNA crosslinking agents. The endogenous function of the FA pathway, however, remains unknown. We have recently demonstrated that the FA pathway is functionally required for the cellular response to disruption of DNA replication. The FA pathway is strongly activated via the mono-ubiquitination of the FANCD2 protein following treatment with the DNA polymerase inhibitor aphidicolin (APH). Furthermore, abrogation of the FA pathway leads to increased chromosome breakage, including breakage at the common chromosomal fragile sites FRA3B and FRA16D, following treatment with APH. We hypothesize that the FA pathway plays an integral role in the maintenance of DNA replication fork stability. Two aims are proposed to gain insight into this function. First, we will characterize the interaction between the FANCD2 protein and the DNA polymerase processivity factor PCNA. Second, we will determine if the increased APH-induced chromosome breakage in FA cells leads to increased integration of foreign DNA. Finally, we will determine if the increased common chromosomal fragile site breakage of FA cells is associated with an increased frequency of human papillomavirus (HPV) DNA integration at these loci, by mapping the genomic sites of HPV integration in FA patient tumors. Our proposed studies will lead to new findings regarding the in vivo physiological role of the FA pathway, and provide insight into the increased cancer susceptibility of FA patients.