A Structural Approach to Unveil the Role of BRCA1 in the Context of Transcription

dc.contributor.authorWinton, Carly Elizabethen
dc.contributor.committeechairKelly, Deborah F.en
dc.contributor.committeememberPoelzing, Stevenen
dc.contributor.committeememberSheng, Zhien
dc.contributor.departmentBiomedical Engineeringen
dc.date.accessioned2016-01-20T09:02:04Zen
dc.date.available2016-01-20T09:02:04Zen
dc.date.issued2016-01-19en
dc.description.abstractThe research presented in this thesis aims to uncover the intricate manner in which BRCA1 interacts with RNAP II during mRNA production utilizing a unique microchip system developed in our lab. We were first able to prove the effectiveness of our tunable system using a breast cancer model of patient derived triple negative breast cancer (TNBC) cells. Here we switched out different mammalian antibodies and collected images of the same structure from different angles. This served many purposes: (1) it proved the system could be tuned for specific uses; (2) it demonstrated that all subunits were present in the complex; (3) it eliminated the need for the tilt function allowing for a less intensive computational procedure. In the BRCA1 wild type cell line we were able to incorporate into our 3D reconstruction, atomic models of the BRCA1-BARD1 heterodimer and the RNAP II core in regions of major unoccupied density. Other areas of minor missing density were overlaid with a short strand of DNA and ubiquitin moieties, which proved agreement with Co-IPs. Next we sought to compare the wild type structure with a BRCA1 mutant variety. Using these techniques, we determined the 3D structure of the mutated complex. After further analysis slight differences were detected between the two complexes, especially in the placement of the atomic models. Overall we were able to determine structural abnormalities that occur when a mutation is present in BRCA1 that may have future applications for targetable therapy in TNBC patients. Moreover, by using TNBC as the disease model, we have created a platform that can be used to evaluate other human diseases due to the tunable nature of our microchip system.en
dc.description.degreeMaster of Scienceen
dc.format.mediumETDen
dc.identifier.othervt_gsexam:6966en
dc.identifier.urihttp://hdl.handle.net/10919/64477en
dc.publisherVirginia Techen
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subjectBreast canceren
dc.subjectBRCA1en
dc.subjectelectron microscopyen
dc.subjecttranscriptionen
dc.titleA Structural Approach to Unveil the Role of BRCA1 in the Context of Transcriptionen
dc.typeThesisen
thesis.degree.disciplineBiomedical Engineeringen
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen
thesis.degree.levelmastersen
thesis.degree.nameMaster of Scienceen

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