title: Human MutS and FANCM complexes function as redundant DNA damage sensors in the Fanconi Anemia pathway.
abstract: The Fanconi Anemia (FA) pathway encodes a DNA damage response activated by DNA damage-stalled replication forks. Current evidence suggests that the FA pathway initiates with DNA damage recognition by the FANCM complex (FANCM/FAAP24/MHF). However, genetic inactivation of FANCM in mouse and DT40 cells causes only a partial defect in the FA pathway activation, suggesting the existence of redundant DNA damage sensors. Here we show that the MutS homologs function in this capacity. A RNAi screen revealed that MSH2 silencing caused defective FA pathway activation, as assessed by damage-induced FANCD2 mono-ubiquitination. A similar FA pathway defect was observed with MSH3 or MSH6 silencing. MSH2 depletion caused cellular phenotypes associated with defective FA pathway, including mitomycin C hypersensitivity and chromosomal instability. Further, silencing of FANCM in MSH2 deficient HEC59 cells caused a more severe FA defect relative to comparable silencing in MSH2 complemented HEC59+Chr2 cells, suggesting redundant functions between MSH2 and FANCM. Consistent with this hypothesis, depletion of MSH2 resulted in defective chromatin localization of the FA core complex upon DNA damage. Further, MSH2 was co-purified and co-immunoprecipitated with FA core complex components. Taken together, our results suggest that human MutS homologs and FANCM complexes function as redundant DNA damage sensors of the FA pathway.
authors.affiliation: Department of Radiation Oncology, Dana-Farber Cancer Institute, 450 Brookline Ave., Boston, MA 02215, United States.
authors.forename: Min
authors.initials: M
authors.lastname: Huang
authors.forename: Richard
authors.initials: R
authors.lastname: Kennedy
authors.forename: Abdullah M
authors.initials: AM
authors.lastname: Ali
authors.forename: Lisa A
authors.initials: LA
authors.lastname: Moreau
authors.forename: Amom Ruhikanta
authors.initials: AR
authors.lastname: Meetei
authors.forename: Alan D
authors.initials: AD
authors.lastname: D'Andrea
authors.forename: Clark C
authors.initials: CC
authors.lastname: Chen
mesh_terms: D000818:Animals; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism; D004249:DNA Damage; D004265:DNA Helicases / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D005199:Fanconi Anemia / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D020868:Gene Silencing; D006801:Humans; D051379:Mice
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; C518295:FANCM protein, human; D004265:DNA Helicases