| Abstract | Our project focused on the conservation of the protein brd-1 in C. elegans. C. elegans is a strong model organism for our study because Ce-brd-1 is the worm ortholog to BARD1 in humans. Specifically, our focus is on its function as an enzyme to attach ubiquitin to the H2A tail of nucleosomes. We studied a structural mutation of Ce-brd-1 that we predicted would interfere with its ability to bind its substrate, the nucleosome. We hypothesized that Ce-brd-1 is bound to the nucleosome at this mutation site based on prior research in the human protein. Therefore, we integrated mutations found in humans into the DNA that codes for C. elegans Ce-brd-1. A typical mutagenesis protocol was used to implement the mutations and then we expressed the proteins in E. coli cells. After that, nucleosomes were reconstituted by dialysis and enzyme activity was assessed using a ubiquitination assay. These assays showed that Ce-brd-1 in C. elegans does bind the nucleosome demonstrating conservation of the BARD1 function. Determining that function is conserved allowed us to determine that C. elegans is an appropriate organism to test mutations found in coserved areas between Ce-brd-1 and BARD1. This research has future clinical potential due to the ability to test mutations encountered in humans using a model organism and can aid with clinical treatment plans to help avoid the development of cancer. |
