title: A Dominant Mutation in Human RAD51 Reveals Its Function in DNA Interstrand Crosslink Repair Independent of Homologous Recombination.
abstract: Repair of DNA interstrand crosslinks requires action of multiple DNA repair pathways, including homologous recombination. Here, we report a de novo heterozygous T131P mutation in RAD51/FANCR, the key recombinase essential for homologous recombination, in a patient with Fanconi anemia-like phenotype. In vitro, RAD51-T131P displays DNA-independent ATPase activity, no DNA pairing capacity, and a co-dominant-negative effect on RAD51 recombinase function. However, the patient cells are homologous recombination proficient due to the low ratio of mutant to wild-type RAD51 in cells. Instead, patient cells are sensitive to crosslinking agents and display hyperphosphorylation of Replication Protein A due to increased activity of DNA2 and WRN at the DNA interstrand crosslinks. Thus, proper RAD51 function is important during DNA interstrand crosslink repair outside of homologous recombination. Our study provides a molecular basis for how RAD51 and its associated factors may operate in a homologous recombination-independent manner to maintain genomic integrity.
authors.affiliation: Laboratory of Genome Maintenance, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Anderson T
authors.initials: AT
authors.lastname: Wang
authors.affiliation: Department of Microbiology and Molecular Genetics, University of California, Davis, Davis, CA 95616, USA.
authors.forename: Taeho
authors.initials: T
authors.lastname: Kim
authors.affiliation: Blood and Marrow Transplant Program, Department of Pediatrics, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: John E
authors.initials: JE
authors.lastname: Wagner
authors.affiliation: Laboratory of Genome Maintenance, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Brooke A
authors.initials: BA
authors.lastname: Conti
authors.affiliation: Laboratory of Genome Maintenance, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Francis P
authors.initials: FP
authors.lastname: Lach
authors.affiliation: Laboratory of Genome Maintenance, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Athena L
authors.initials: AL
authors.lastname: Huang
authors.affiliation: Proteomics Resource Center, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Henrik
authors.initials: H
authors.lastname: Molina
authors.affiliation: Laboratory of Genome Maintenance, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Erica M
authors.initials: EM
authors.lastname: Sanborn
authors.affiliation: Blood and Marrow Transplant Program, Department of Pediatrics, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Heather
authors.initials: H
authors.lastname: Zierhut
authors.affiliation: New York Genome Center, New York, NY 10013, USA.
authors.forename: Belinda K
authors.initials: BK
authors.lastname: Cornes
authors.affiliation: New York Genome Center, New York, NY 10013, USA.
authors.forename: Avinash
authors.initials: A
authors.lastname: Abhyankar
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
authors.forename: Carrie
authors.initials: C
authors.lastname: Sougnez
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
authors.forename: Stacey B
authors.initials: SB
authors.lastname: Gabriel
authors.affiliation: Human Genetics and Hematology, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Arleen D
authors.initials: AD
authors.lastname: Auerbach
authors.affiliation: Department of Microbiology and Molecular Genetics, University of California, Davis, Davis, CA 95616, USA.
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Kowalczykowski
authors.affiliation: Laboratory of Genome Maintenance, The Rockefeller University, New York, NY 10065, USA. Electronic address: asmogorzewska@rockefeller.edu.
authors.forename: Agata
authors.initials: A
authors.lastname: Smogorzewska
mesh_terms: D002470:Cell Survival; D003432:Cross-Linking Reagents; D004247:DNA / Q000378:metabolism*; D004265:DNA Helicases / Q000378:metabolism; D004260:DNA Repair; D005090:Exodeoxyribonucleases / Q000378:metabolism; D005199:Fanconi Anemia / Q000235:genetics* / Q000378:metabolism; D005260:Female; D042822:Genomic Instability; D057809:HEK293 Cells; D006579:Heterozygote; D006801:Humans; D007223:Infant; D009154:Mutation; D051135:Rad51 Recombinase / Q000235:genetics* / Q000378:metabolism*; D053484:RecQ Helicases / Q000378:metabolism; D051716:Replication Protein A / Q000378:metabolism*; D000071657:Werner Syndrome Helicase
chemical_list: D003432:Cross-Linking Reagents; C497167:RPA1 protein, human; D051716:Replication Protein A; D004247:DNA; C495153:RAD51 protein, human; D051135:Rad51 Recombinase; D005090:Exodeoxyribonucleases; D004265:DNA Helicases; C509908:DNA2 protein, human; D053484:RecQ Helicases; C098785:WRN protein, human; D000071657:Werner Syndrome Helicase