The heterotrimeric G proteins regulate a remarkable breadth of biological processes, from maintenance of homeostasis to perception of the environment to development of the organism. In this proposal, we will test the hypothesis that combinatorial association of the alpha, beta and gamma subunits comprising the G proteins may provide a mechanism for the propagation of such a wide range of biological responses. For this purpose, zebrafish offer a number of specific advantages. Because they exhibit a similar molecular complexity to humans and mice, zebrafish provide an unique system to study the ontogeny of this important class of multi-subunit proteins. Whole mount in situ hybridization will be used to identify when and where the various alpha, beta, and gamma subtypes are expressed. Subsequently, loss-of-function analysis using a morpholino anti-sense strategy will be used to directly link a specific subtype to a particular function and signaling pathway. Finally, complementation analysis using an expression approach to try to rescue the loss-of-function phenotype will be used to determine what level of functional redundancy exists among the related subtypes. This information will provide a framework for understanding how the G proteins are able to receive, integrate, and process the multitude of signals required for the development of the organism. Since many diseases (e.g. cancer, heart disease) are associated with re-induction of embryonic genes, this information will eventually lead to the better diagnosis and more selective therapeutic treatments.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM058191-06A2
Application #
6776055
Study Section
Pharmacology A Study Section (PHRA)
Program Officer
Lograsso, Philip
Project Start
1998-09-01
Project End
2008-03-31
Budget Start
2004-04-01
Budget End
2005-03-31
Support Year
6
Fiscal Year
2004
Total Cost
$281,992
Indirect Cost
Name
Weis Center for Research-Geisinger Clinc
Department
Type
DUNS #
079161360
City
Danville
State
PA
Country
United States
Zip Code
17822
Leung, Tinchung; Humbert, Jasper E; Stauffer, Anna M et al. (2008) The orphan G protein-coupled receptor 161 is required for left-right patterning. Dev Biol 323:31-40
Chen, Hui; Leung, Tinchung; Giger, Kathryn E et al. (2007) Expression of the G protein gammaT1 subunit during zebrafish development. Gene Expr Patterns 7:574-83
Leung, Tinchung; Chen, Hui; Stauffer, Anna M et al. (2006) Zebrafish G protein gamma2 is required for VEGF signaling during angiogenesis. Blood 108:160-6
Sudol, Marius; Recinos, Claudia C; Abraczinskas, Jennifer et al. (2005) WW or WoW: the WW domains in a union of bliss. IUBMB Life 57:773-8
Robishaw, Janet D; Berlot, Catherine H (2004) Translating G protein subunit diversity into functional specificity. Curr Opin Cell Biol 16:206-9
Cheng, Keith C; Levenson, Robert; Robishaw, Janet D (2003) Functional genomic dissection of multimeric protein families in zebrafish. Dev Dyn 228:555-67
Robishaw, Janet D; Wang, Qin; Schwindinger, William F (2002) Ribozyme-mediated suppression of G protein gamma subunits. Methods Enzymol 344:435-51
Wang, Q; Jolly, J P; Surmeier, J D et al. (2001) Differential dependence of the D1 and D5 dopamine receptors on the G protein gamma 7 subunit for activation of adenylylcyclase. J Biol Chem 276:39386-93
Wang, Q; Mullah, B K; Robishaw, J D (1999) Ribozyme approach identifies a functional association between the G protein beta1gamma7 subunits in the beta-adrenergic receptor signaling pathway. J Biol Chem 274:17365-71