RUI: Functional Analysis of the C. elegans Winged-Helix Transcription Factor, LIN-31

Santa Clara University, Santa Clara CA

Investigators

Abstract

0315072 Miller The LIN-31 protein, a member of the winged-helix family of transcription factors, is required for the proper specification of vulval cell fates during C. elegans development. This protein is believed to play two roles during development: (1) to promote vulval cell fates (when phosphorylated by the signal transduction protein MAP kinase) and (2) to promote non-vulval cell fates (when heterodimerized with another transcription factor, LIN-1). This project aims to further investigate LIN-31's role in cell fate specification during vulval development by continuing a functional analysis of this multifaceted protein. The Specific Aims of this proposal are (1) Functional Analysis of the Winged Helix Transcription Factor LIN-31 (site-directed mutagenesis experiments will assign functions to different regions of the LIN-31 protein), (2) Identification of LIN- 31::LIN-1 Heterodimerization Elements (co-immunoprecipitation experiments will be used to determine if the acidic region of the LIN-31 protein is required for heterodimerization with the LIN-1 protein, a known heterodimerization partner), and (3) Identification of High-Affinity LIN-31 Binding Sequences (a sequential selection technique will be used to identify oligonucleotides that bind the LIN-31 DNA binding domain with high affinity--those sequences will then be used to identify actual targets of the LIN-31 transcription factor). The Intellectual Merit of this project is to provide a better understanding of how gene regulation controls development by advancing our knowledge of a transcription factor at the end of a signaling cascade that determines cell fates. In addition, by identifying potential targets of this transcription factor, the experiments described in this proposal will also illuminate a part of the C. elegans vulval cell fate pathway that is not well understood. The Broader Impacts of this project include significant contributions to teaching, training and learning. Undergraduate students will be an integral part of the project, involvement of the P.I. in research will enhance her teaching, and training of a Teaching Post-Doc will provide critical training in education and research, in addition to enhancing the evolving "community of science scholars" at SCU. Furthermore, of the 23 undergraduate students who have thus far conducted research in the P.I.'s laboratory, 17 (74%) were women, most of which have gone on to post-graduate programs. These numbers indicate established success in recruiting and educating members of underrepresented groups. SCU also has a significant Latino population, from which research students will continue to be recruited.