The overall goal of this project is to understand how a novel adaptor protein and Src substrate, Fish, contributes to tumorigenic transformation. Src was the first oncogene to be discovered, and the first tyrosine kinase. Src is a very potent oncogene. It transforms cells by stimulating cell division, by eliciting the production of autocrine growth factors, by preventing apoptosis, and by increasing cell motility and invasiveness. Each of these pathways is important to understand since they are also features of human tumors. Indeed, Src activity has been shown to be elevated in human tumors, particularly those of breast and colon, and its increased activity has been correlated with a more aggressive phenotype. Although we know much of how Src switches on signal transduction pathways, we cannot yet fully describe how it participates in pathways controlling growth factor production, movement and invasion. This is likely because of an incomplete description of Src substrates. We have isolated a novel substrate of Src called Fish. Fish is a scaffold, or adaptor, protein. That is, it lacks catalytic activity, but contains multiple motifs that allow it to associate with other proteins, as well as with lipids. Fish has one PX domain, five SH3 domains, as well as multiple motifs that could mediate association with SH2- and SH3-domain containing proteins. We have recently found that Fish associates with the cytoplasmic tails of members of the ADAMs family. These are proteins with metalloprotease, adhesion and fusion domains, that are involved in shedding of active growth factors from cells, ceil-cell interactions, and fusion of specialized cells such as myoblasts. Some members of the ADAMs family are over-expressed in human cancers. Our long term objective is to understand how phosphorylation of Fish by Src, and association of Fish with ADAMs family proteins (and other signaling molecules) contributes to normal and tumor cell physiology. We propose to characterize the interaction of Fish with the ADAMs in more detail as well as isolate more Fish-interacting proteins. We will also define Fish function in development by creating and analyzing Fish knockout mice. And finally, we will test the hypothesis that the Fish-ADAM association mediates the production of autocrine growth factors, and/or motility, observed in both Src-transformed cells and human tumor cells. These studies will contribute to our understanding of the mechanisms of oncogenesis.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA098383-06
Application #
7339325
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Jhappan, Chamelli
Project Start
2004-02-19
Project End
2010-01-31
Budget Start
2008-02-01
Budget End
2010-01-31
Support Year
6
Fiscal Year
2008
Total Cost
$334,133
Indirect Cost
Name
Sanford-Burnham Medical Research Institute
Department
Type
DUNS #
020520466
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Cejudo-Martin, Pilar; Yuen, Angela; Vlahovich, Nicole et al. (2014) Genetic disruption of the sh3pxd2a gene reveals an essential role in mouse development and the existence of a novel isoform of tks5. PLoS One 9:e107674
Courtneidge, Sara A (2012) Cell migration and invasion in human disease: the Tks adaptor proteins. Biochem Soc Trans 40:129-32
Murphy, Danielle A; Courtneidge, Sara A (2011) The 'ins' and 'outs' of podosomes and invadopodia: characteristics, formation and function. Nat Rev Mol Cell Biol 12:413-26
Quintavalle, Manuela; Elia, Leonardo; Condorelli, Gianluigi et al. (2010) MicroRNA control of podosome formation in vascular smooth muscle cells in vivo and in vitro. J Cell Biol 189:13-22
Iqbal, Zafar; Cejudo-Martin, Pilar; de Brouwer, Arjan et al. (2010) Disruption of the podosome adaptor protein TKS4 (SH3PXD2B) causes the skeletal dysplasia, eye, and cardiac abnormalities of Frank-Ter Haar Syndrome. Am J Hum Genet 86:254-61
Stylli, Stanley S; Stacey, T T I; Verhagen, Anne M et al. (2009) Nck adaptor proteins link Tks5 to invadopodia actin regulation and ECM degradation. J Cell Sci 122:2727-40
Diaz, Begoña; Shani, Gidon; Pass, Ian et al. (2009) Tks5-dependent, nox-mediated generation of reactive oxygen species is necessary for invadopodia formation. Sci Signal 2:ra53
Crimaldi, Luca; Courtneidge, Sara A; Gimona, Mario (2009) Tks5 recruits AFAP-110, p190RhoGAP, and cortactin for podosome formation. Exp Cell Res 315:2581-92
Gianni, Davide; Diaz, Begoña; Taulet, Nicolas et al. (2009) Novel p47(phox)-related organizers regulate localized NADPH oxidase 1 (Nox1) activity. Sci Signal 2:ra54
Blouw, Barbara; Seals, Darren F; Pass, Ian et al. (2008) A role for the podosome/invadopodia scaffold protein Tks5 in tumor growth in vivo. Eur J Cell Biol 87:555-67