Mary Bello
Research Mentor(s): Jacob Mueller
Department or Program: Human Genetics
Authors: Mary Bello, Ann Marie, Jacob Mueller
Session: Session 2: 1:00pm-1:50pm
Poster: 3
Abstract
Many genes are conserved throughout multiple species, and retain the same function in the same tissues. However, some genes are duplicated and these duplicates can acquire novel functions. The nuclear RNA export factor 1 (Nxf1) gene is a highly conserved autosomal gene essential for life in eukaryotes. Nxf1 exports nuclear RNA to the cytoplasm, but in mammals it also has X-linked gene duplicates. One of these Nxf1 duplicates, nuclear RNA export factor 2 (Nxf2), has an unknown function in mice. Nxf2 is expressed in the germ cells of the testis, and is required for male fertility. Similar to Nxf1’s essential cofactor Nxt1, Nxf2 is also known to interact with a cofactor called Nxt2. In order to test whether Nxf2 has acquired a novel function in the male germline compared to Nxf1, we will clone the Nxf1, Nxf2, Nxt1, and Nxt2 genes into yeast expression vectors. These expression vectors can then be used to perform a series of yeast 2-hybrid assays which will test whether different combinations of the previously mentioned proteins are capable of interacting. Our expected results are that NXF1 and NXT1 as well as NXF2 and NXT2 will interact, and therefore will serve as positive controls. We will also test the capability of NXF2 to interact with NXT1. If NXF2, like NXF1, binds NXT1, this would suggest NXF1 and NXF2 may perform the same function. However, if NXF2 does not interact with NXT1, it is likely because it performs a different function than NXF1. Additionally, our lab has preliminary data that NXF2 interacts with RNA-binding protein 44 (RBM44), a protein known to localize to the intercellular bridges connecting the cytoplasm of male germ cells during spermatogenesis. The results from these yeast 2-hybrid assays will provide information on what proteins NXF2 binds, and whether they are similar or different to those NXF1 binds. Evidence for NXF2 binding different proteins from NXF1 would support that the Nxf2 gene has evolved a drastically different function from its progenitor Nxf1, and this role is potentially restricted to the cytoplasm.
