Effect of mutations in BRCA1 tumor suppressor protein on BRCA1-DNA interactions
This project is a part of our effort to build a coherent mechanistic description of molecular events that ensure accurate DNA repair by homologous recombination. The target of this project is BRCA1 tumor suppressor protein. We are applying traditional biochemical analyses and our novel single-molecule sorting methodology to establish robust and inexpensive functional assays, which will allow correlating pathogenic BRCA1 defects with BRCA1 mutations and/or with the presence of a particular array of posttranslational modifications affecting BRCA1 activities and interactions. Such functional assays will be instrumental in predicting whether a particular mutation in BRCA1 gene or the array of PTMs found in a patient are predictive of cancer predisposition or purely neutral. Recently, we established a single-molecule assay to monitor interaction between the DNA binding region of BRCA1 with various DNA structures. This assay will be used to probe the effects of mutations found in breast cancer patients on BRCA1-DNA interactions. Assigning functional significance to numerous uncharacterized variants of BRCA1 will allow the carriers of disease-causing mutations to be identified during genetic testing.
basic molecular biology skills; university level math, physics, chemistry; if taken biochemistry/physical chemistry/biophysics courses will give some advantage, but are not required
Based on the information on the missense variants or in-frame deletions in the BRCA1 are reported in the Breast Cancer Information Core database, we will select the most commonly occurring uncharacterized variants of BRCA1 gene. Using site-directed mutagenesis, the student will produce mutant forms of BRCA1 DNA binding regions and purify corresponding proteins. The effect of the mutations on BRCA1-DNA binding will be accessed using our single-molecule fluorescence based assays.