title: A genetic screen identifies FAN1, a Fanconi anemia-associated nuclease necessary for DNA interstrand crosslink repair.
abstract: The Fanconi anemia (FA) pathway is responsible for interstrand crosslink repair. At the heart of this pathway is the FANCI-FAND2 (ID) complex, which, upon ubiquitination by the FA core complex, travels to sites of damage to coordinate repair that includes nucleolytic modification of the DNA surrounding the lesion and translesion synthesis. How the ID complex regulates these events is unknown. Here we describe a shRNA screen that led to the identification of two nucleases necessary for crosslink repair, FAN1 (KIAA1018) and EXDL2. FAN1 colocalizes at sites of DNA damage with the ID complex in a manner dependent on FAN1's ubiquitin-binding domain (UBZ), the ID complex, and monoubiquitination of FANCD2. FAN1 possesses intrinsic 5'-3' exonuclease activity and endonuclease activity that cleaves nicked and branched structures. We propose that FAN1 is a repair nuclease that is recruited to sites of crosslink damage in part through binding the ubiquitinated ID complex through its UBZ domain.
authors.affiliation: Howard Hughes Medical Institute, Department of Genetics, Harvard Medical School, and Brigham and Women's Hospital, Boston, MA 02115, USA. asmogorzewska@rockefeller.edu
authors.forename: Agata
authors.initials: A
authors.lastname: Smogorzewska
authors.forename: Rohini
authors.initials: R
authors.lastname: Desetty
authors.forename: Takamune T
authors.initials: TT
authors.lastname: Saito
authors.forename: Michael
authors.initials: M
authors.lastname: Schlabach
authors.forename: Francis P
authors.initials: FP
authors.lastname: Lach
authors.forename: Mathew E
authors.initials: ME
authors.lastname: Sowa
authors.forename: Alan B
authors.initials: AB
authors.lastname: Clark
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Kunkel
authors.forename: J Wade
authors.initials: JW
authors.lastname: Harper
authors.forename: Monica P
authors.initials: MP
authors.lastname: Colaiácovo
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Elledge
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans / Q000378:metabolism; D002460:Cell Line; D003432:Cross-Linking Reagents / Q000378:metabolism*; D004249:DNA Damage; D053843:DNA Mismatch Repair / Q000187:drug effects; D004260:DNA Repair / Q000187:drug effects; D004706:Endodeoxyribonucleases; D004720:Endonucleases / Q000378:metabolism; D005090:Exodeoxyribonucleases / Q000737:chemistry / Q000378:metabolism*; D005092:Exonucleases / Q000737:chemistry / Q000378:metabolism*; D005199:Fanconi Anemia / Q000201:enzymology* / Q000473:pathology; D052236:Fanconi Anemia Complementation Group D2 Protein / Q000378:metabolism; D005820:Genetic Testing / Q000379:methods*; D015894:Genome, Human / Q000235:genetics; D006801:Humans; D016685:Mitomycin / Q000494:pharmacology; D008969:Molecular Sequence Data; D064251:Multifunctional Enzymes; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000187:drug effects; D034741:RNA, Small Interfering / Q000378:metabolism
chemical_list: D003432:Cross-Linking Reagents; D052236:Fanconi Anemia Complementation Group D2 Protein; D064251:Multifunctional Enzymes; D034741:RNA, Small Interfering; D016685:Mitomycin; D004706:Endodeoxyribonucleases; D004720:Endonucleases; D005090:Exodeoxyribonucleases; D005092:Exonucleases; C551063:FAN1 protein, human