We will make fully operational a continuing pilot collaboration between the labs of Dr. Jessica P. Houston (NMSU) and Dr. Roger Brent (FHCRC). Our proposed work builds on complementary strengths in the two labs: instrumentation engineering and signal analysis in the Houston lab, protein engineering and genetics in the Brent lab. In particular, the work builds on "frequency domain" methods to determine fluorescence lifetimes in microscopy, and on significantly more advanced developments in "frequency domain" methods for flow cytometry, for which Dr. Houston is emerging as a leader. Dysfunctions in cell signaling leading to inappropriate cell proliferation contribute to most cancers. Although much is known about cell signaling, key questions, including the causes and consequences of cell-to-cell variation in signaling and response, remain unanswered. Here, we will use fluorescent lifetime methods to extend the power of single cell assays to quantify key events in cell signaling in yeast (the "development platform") and in mammalian cells. These measurements depend on the use of Green Fluorescent Protein (GFP) derivatives fused to the proteins that make up the different cell signaling pathways to quantify events such as recruitment of a signaling complex to the cell membrane. During the next three years, we will develop our methods, to address these questions and to allow wider applications. In particular, we will continue to improve our instrumentation, use it to develop better GFP derivatives and signaling reporters, and use the improved methods to understand the causes of cell-to-cell variation in a yeast signaling system. We will also use these methods to develop high-signal reporters that allow quantification of cell signaling in clonal, mammalian cell lines using flow cytometry. Successful work will merit future funding. It should also increase the visibility of the Houston lab and help recruit new students and researchers from underrepresented ethnic minorities into an active area of international scientific research.

National Institute of Health (NIH)
Specialized Center--Cooperative Agreements (U54)
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Special Emphasis Panel (ZCA1)
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Fred Hutchinson Cancer Research Center
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O'Keefe, David D; Thomas, Sean; Edgar, Bruce A et al. (2014) Temporal regulation of Dpp signaling output in the Drosophila wing. Dev Dyn 243:818-32
Song, Mingzhou; Zhang, Yang; Katzaroff, Alexia J et al. (2014) Hunting complex differential gene interaction patterns across molecular contexts. Nucleic Acids Res 42:e57
Thompson, Beti; Vilchis, Hugo; Moran, Crystal et al. (2014) Increasing cervical cancer screening in the United States-Mexico border region. J Rural Health 30:196-205
Sands, Bryan; Jenkins, Patrick; Peria, William J et al. (2014) Measuring and sorting cell populations expressing isospectral fluorescent proteins with different fluorescence lifetimes. PLoS One 9:e109940
Thompson, Beti; O'Connell, Mary; Loest, Helena et al. (2013) Understanding and reducing obstacles in a collaboration between a minority institution and a cancer center. J Health Care Poor Underserved 24:1648-56
Coronado, Gloria D; Ondelacy, Stephanie; Schwarz, Yvonne et al. (2012) Recruiting underrepresented groups into the Carbohydrates and Related Biomarkers (CARB) cancer prevention feeding study. Contemp Clin Trials 33:641-6
O'Keefe, David D; Edgar, Bruce A; Saucedo, Leslie J (2011) EndoGI modulates Notch signaling and axon guidance in Drosophila. Mech Dev 128:59-70
Coronado, Gloria D; O'Connell, Mary A; Anderson, Jennifer et al. (2010) Undergraduate cancer training program for underrepresented students: findings from a minority institution/cancer center partnership. J Cancer Educ 25:32-5
Metzger, Michael J; Holguin, Christiana J; Mendoza, Ramon et al. (2010) The prostate cancer-associated human retrovirus XMRV lacks direct transforming activity but can induce low rates of transformation in cultured cells. J Virol 84:1874-80
Thompson, Beti; Ondelacy, Stephanie; Godina, Ruby et al. (2010) A small grants program to involve communities in research. J Community Health 35:294-301

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