Intellectual Merit. Notch signaling is an important and highly significant topic to understand because it represents one of the five major signal transduction pathways (Wnt/beta-catenin, Hedgehog, receptor tyrosine kinases, nuclear receptors, and Notch) found in all metazoan organisms responsible for growth, differentiation, and maintenance of cells. Misregulation of Notch signaling can result in developmental as well as disease states. After ligand activation of the Notch receptor, an intracellular segment (NICD) translocates to the nucleus. The NICD forms a ternary complex with the Mam (Mastermind) protein and other proteins to activate target genes. In humans, there are three Mam proteins (Mam1-3). Each Mam contains an amino terminal basic domain that interacts with NICD and a carboxy terminal acidic region that likely functions in transcriptional activation. Truncated versions of Mam1 that lose these carboxyl regions, but retain the basic domain, produce dominant negative phenotypes when they are over-expressed in model organisms. The overall goal of this project is to dissect the molecular elements of the Mam family of proteins that are necessary for Notch signaling with the goal of gaining insight into the complexity at the molecular level. Yeast two-hybrid screens and biochemical protein-protein pull down techniques will be employed to identify Mam-interacting partners from tissue culture cell lines. The role of putative Mam-interacting proteins will be determined though a combination of molecular and biochemical approaches.
Broader Impacts. This project will benefit undergraduate students who will receive training in biochemistry and molecular biology at San José State University (SJSU). The majority of students attending SJSU are from economically disadvantaged families. A number of these students are from underrepresented or first time college attendees. This project will provide students with a unique opportunity to learn and conduct research. Students will be encouraged to think and design their own experiments and will present their results at both state and national conferences. Further, to reach as many students as possible, some of the research methods developed during this project will also be incorporated into an undergraduate molecular biology laboratory course.
