NSF Award
9252959
The project has two objectives: (1) for the investigator to learn molecular cloning by isolating a new member of the TGF- beta superfamily from mammalian cells through work in the host laboratory, learning state-of-the-art cloning of membrane proteins; and (2) to make sea urchin cDNA libraries from embryos, and with these and host laboratory resources, to search for clones for TFG- beta receptors and integrin receptors. The investigator will begin by using low-stringency hybridization to look for sea urchin clones of TGF-beta and integrin receptors. If too many clones are found she will design redundant oligo-nucleotide probes and use PCR techniques to try to isolate sea urchin probes from her libraries, or prepare antibodies to redundant peptide sequences in the integrin receptors and screen expression libraries for clones. Finally, she will use her own antibodies to the putative 200kDa-laminin receptor to screen for clones in the sea urchin libraries. The long term goal is to understand how receptor molecules on the surface of cells interact with specific molecules of the extracellular matrix to signal or aid cell movements and morphogenesis of embryos. With preliminary evidence that the interaction of the extracellular matrix protein, laminin, by interacting with cell receptors may stimulate or mediate gastrulation these embryos, the investigator has begun to purify membrane proteins from sea urchin embryos that bind to extracellular matrix components. She has found one protein that binds laminin--the first identification of a matrix receptor in sea urchin embryos, and is currently searching for other integrin proteins that may bind laminin or fibronectin using an antibody evolutionary tree from sea urchins to mammals, significantly overlap. The investigator, an expert on the regulation of translation in echinoderm egg and embryos, is changing the emphasis of her own laboratory to study the role of membrane receptors and extracellular matrix molecules in morphogenetic movements of sea urchin embryos. The studies on sea urchins, a model for the easy visualization and manipulation of development, are potentially very important for furthering our understanding of cell-extracellular matrix interactions as determinants of embryonic cell movement and differentiation, particularly the critical processes of gastrulation, and similar processes such as imaginal disc formation in Drosophila or neurulation in vertebrates. This project furthers VPW program objectives to provide opportunities for women to advance their careers in science or engineering through research, and to encourage other women to pursue careers in these areas through the investigator's enhanced visibility as a role model on the host campus. The proposed activities which contribute to the second objective include: giving lectures/seminars; lecturing in a number of graduate and undergraduate courses, including developmental biology, introductory biology, and advanced cell biology; working with graduate student women participating in the on-going biology brown-bag lunches; and giving a lecture to the Cambridge Chapter of the Association of Women in Cell Biology (AWIS).
