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dc.contributor.advisorElledge, Stephen J.
dc.contributor.authorAdamson, Brittany Susan
dc.date.accessioned2013-03-20T13:06:29Z
dc.date.issued2013-03-20
dc.date.submitted2012
dc.identifier.citationAdamson, Brittany Susan. 2012. A Genome-Wide Study of Homologous Recombination in Mammalian Cells Identifies RBMX, a Novel Component of the DNA Damage Response. Doctoral dissertation, Harvard University.en_US
dc.identifier.otherhttp://dissertations.umi.com/gsas.harvard:10723en
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:10448771
dc.description.abstractRepair of DNA double-strand breaks is critical to the maintenance of genomic stability, and failure to repair these DNA lesions can cause loss of chromosome telomeric regions, complex translocations, or cell death. In humans this can lead to severe developmental abnormalities and cancer. A central pathway for double-strand break repair is homologous recombination (HR), a mechanism that operates during the S and G2 phases of the cell cycle and primarily utilizes the replicated sister chromatid as a template for repair. Most knowledge of HR is derived from work carried out in prokaryotic and eukaryotic model organisms. To probe the HR pathway in human cells, we performed a genome-wide siRNA-based screen; and through this screen, we uncovered cellular functions required for HR and identified proteins that localize to sites of DNA damage. Among positive regulators of HR, we identified networks of pre-mRNA-processing factors and canonical DNA damage response effectors. Within the former, we found RBMX, a heterogeneous nuclear ribonucleoprotein (hnRNP) that associates with the spliceosome, binds RNA, and influences alternative splicing. We found that RBMX is required for cellular resistance to genotoxic stress, accumulates at sites of DNA damage in a poly(ADP-ribose) polymerase 1-dependent manner and through multiple domains, and promotes HR by facilitating proper BRCA2 expression. Screen data also revealed that the mammalian recombinase RAD51 is commonly off-targeted by siRNAs, presenting a cautionary note to those studying HR with RNAi and highlighting the vulnerability of RNAi screens to off-target effects in general. Candidate validation through secondary screening with independent reagents successfully circumvented the effects of off-targeting and set a new standard for reagent redundancy in RNAi screens.en_US
dc.language.isoen_USen_US
dash.licenseLAA
dc.subjectGeneticsen_US
dc.subjecthomologous recombinationen_US
dc.subjectOff-targeten_US
dc.subjectRAD51en_US
dc.subjectRBMXen_US
dc.subjectRNAien_US
dc.subjectscreenen_US
dc.titleA Genome-Wide Study of Homologous Recombination in Mammalian Cells Identifies RBMX, a Novel Component of the DNA Damage Responseen_US
dc.typeThesis or Dissertationen_US
dash.depositing.authorAdamson, Brittany Susan
dc.date.available2013-03-20T13:06:29Z
thesis.degree.date2012en_US
thesis.degree.disciplineGenetics and Genomicsen_US
thesis.degree.grantorHarvard Universityen_US
thesis.degree.leveldoctoralen_US
thesis.degree.namePh.D.en_US
dc.contributor.committeeMemberAmon, Angelikaen_US
dc.contributor.committeeMemberLahav, Galiten_US
dc.contributor.committeeMemberWalter, Johannesen_US
dc.contributor.committeeMemberScully, Ralphen_US
dash.contributor.affiliatedAdamson, Brittany Susan


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