The long-term objectives of this proposal are to identify and characterize proteins that function in signal transduction pathways and discover how these proteins act together to regulate cell fates during development. This information will be important for understanding fundamental aspects of animal development and developing new diagnostic and therapeutic strategies for human diseases that result from abnormalities of signaling pathways, such as birth defects and cancer. The proposed research focuses on an evolutionarily conserved signaling pathway that includes a receptor tyrosine kinase, Ras, mitogen activated protein (MAP) kinase, and ETS transcription factors. These pathways mediate many different cell fate decisions during development, and mutations that affect these proteins are a common cause of human tumors. MAP kinase function will be investigated by characterizing two docking sites that enable MAP kinases to interact with substrate proteins, identifying additional docking sites, determining how docking sites affect the selection of phosphorylation sites in substrate proteins, and identifying the region of MAP kinase that interacts with docking sites. Information about docking sites will be used to investigate how phosphorylation affects protein function, develop peptide inhibitors of MAP kinase and predict new substrates of MAP kinase. Candidate substrates will be evaluated in vitro and in vivo. These studies should illuminate how MAP kinase activity causes different cell fates in different developmental contexts. ETS transcription factors are conserved mediators of MAP kinase signaling, including the LIN-1 protein in Caenorhabditis elegans. LIN-1 regulation will be investigated by characterizing DNA binding, transcriptional regulation, and protein stability, and determining how these properties are regulated by MAP kinase activity. Target genes that are regulated by LIN-1 will be identified using molecular and genetic approaches, and a network of genes that interact with LIN-1 will be identified and characterized. These studies will illuminate how MAP kinases identify and interact with substrate proteins, how substrate proteins mediate different cell fates during animal development, how ETS transcription factors are regulated by MAP kinase and how ETS proteins regulate cell fates during development.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA084271-02
Application #
6342212
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Spalholz, Barbara A
Project Start
2000-01-01
Project End
2004-12-31
Budget Start
2001-01-01
Budget End
2001-12-31
Support Year
2
Fiscal Year
2001
Total Cost
$256,463
Indirect Cost
Name
Washington University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Leight, Elizabeth R; Murphy, John T; Fantz, Douglas A et al. (2015) Conversion of the LIN-1 ETS protein of Caenorhabditis elegans from a SUMOylated transcriptional repressor to a phosphorylated transcriptional activator. Genetics 199:761-75
Roh, Hyun Cheol; Collier, Sara; Deshmukh, Krupa et al. (2013) ttm-1 encodes CDF transporters that excrete zinc from intestinal cells of C. elegans and act in a parallel negative feedback circuit that promotes homeostasis. PLoS Genet 9:e1003522
Roh, Hyun Cheol; Collier, Sara; Guthrie, James et al. (2012) Lysosome-related organelles in intestinal cells are a zinc storage site in C. elegans. Cell Metab 15:88-99
Murphy, John T; Bruinsma, Janelle J; Schneider, Daniel L et al. (2011) Histidine protects against zinc and nickel toxicity in Caenorhabditis elegans. PLoS Genet 7:e1002013
Liu, Zhenyi; Schneider, Daniel L; Kornfeld, Kerry et al. (2010) Simple copy number determination with reference query pyrosequencing (RQPS). Cold Spring Harb Protoc 2010:pdb.prot5491
Johnson, Kenneth G; Kornfeld, Kerry (2010) The CRAL/TRIO and GOLD domain protein TAP-1 regulates RAF-1 activation. Dev Biol 341:464-71
Bruinsma, Janelle J; Schneider, Daniel L; Davis, Diana E et al. (2008) Identification of mutations in Caenorhabditis elegans that cause resistance to high levels of dietary zinc and analysis using a genomewide map of single nucleotide polymorphisms scored by pyrosequencing. Genetics 179:811-28
Leight, Elizabeth R; Glossip, Danielle; Kornfeld, Kerry (2005) Sumoylation of LIN-1 promotes transcriptional repression and inhibition of vulval cell fates. Development 132:1047-56
Miley, Ginger R; Fantz, Douglas; Glossip, Danielle et al. (2004) Identification of residues of the Caenorhabditis elegans LIN-1 ETS domain that are necessary for DNA binding and regulation of vulval cell fates. Genetics 167:1697-709
Hsu, Virginia; Zobel, Cheri L; Lambie, Eric J et al. (2002) Caenorhabditis elegans lin-45 raf is essential for larval viability, fertility and the induction of vulval cell fates. Genetics 160:481-92

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