Hedgehog Signaling And Intracellular Transport
Jonathan T Eggenschwiler
Molecular Biologyprinceton University
office Of Research And Project Administration
princeton, Nj 085440036
Grant 5R01HD050761-04 from Eunice Kennedy Shriver National Institute Of Child Health & Human Development IRG: DEV
Abstract: Embryonic cells use the Hedgehog signaling pathway to convey information that controls cell fate, proliferation, and survival. Misregulation of this pathway leads to birth defects and a variety of cancers in humans. Recent data point to key mechanisms controlling the Hedgehog pathway in mammals that do not appear to be evolutionary conserved in invertebrate model organisms. The primary objective of the proposed research is to understand how Hedgehog signal transduction functions at the mechanistic level using the mouse as a model system. Specifically, the roles of 2 novel antagonists of the mammalian pathway, Wdr10 and Tulp3, will be studied using genetic, cellular, and biochemical approaches outlined in 3 specific aims. The Wdr10 protein, as well as its invertebrate homologs, functions in intraflagellar transport, a process used for building cilia and flagella. The experiments proposed in Aim 1 will confirm the requirement for Wdr10 in regulating the Hedgehog pathway through targeted mutagenesis and transgenic rescue approaches. Other experiments will address the subcellular localization of Wdr10 and test whether it acts in retrograde transport using immunocytochemistry and live cell imaging techniques. The mechanism of Wdr10 function in the Hedgehog pathway will be addressed in Aim 2. The hypothesis that Wdr10 acts cell autonomously will be tested through chimera analysis and the step in the Hedgehog pathway at which Wdr10 acts will be clarified through epistasis analysis. In addition, the hypothesis that Wdr10 acts directly or indirectly in controlling nuclear localization of the Gli transcription factors will be pursued using immunohistochemical and biochemical methods. Finally, the experiments in Aim 3 will test 2 hypotheses regarding the function of Tulp3 in Hedgehog signaling. First, the importance of the interaction between Tulp3 and myosin Vb will be functionally tested by blocking myosin Vb activity in vivo. Second, the significance of regulated Tulp3 nuclear translocation in Hedgehog signaling will be examined in tissue culture cells
Keywords: biological signal transduction, genetic regulation, inhibitor /antagonist, intracellular transport, protein structure function, transcription factor, transport protein flagellum, gene mutation, kinesin, myosin, neuronal transport, protein localization, protein protein interaction cell line, genetically modified animal, immunocytochemistry, laboratory mouse, laboratory rabbit, molecular /cellular imaging
Project start date: 2005-07-15
Project end date: 2010-06-30
5R01HD050761-04 (2008): $281286
Sponsored Links Excellgen http://Excellgen.com
HEDGEHOG SIGNALING AND INTRACELLULAR TRANSPORT
Jonathan T Eggenschwiler, Assistant Professor
Princeton University, Office Of Research And Project Administration, Princeton, Nj 08544-0036
Grant 5R01HD050761-05 from Eunice Kennedy Shriver National Institute Of Child Health & Human Development
Abstract: Embryonic cells use the Hedgehog signaling pathway to convey information that controls cell fate, proliferation, and survival. Misregulation of this pathway leads to birth defects and a variety of cancers in humans. Recent data point to key mechanisms controlling the Hedgehog pathway in mammals that do not appear to be evolutionary conserved in invertebrate model organisms. The primary objective of the proposed research is to understand how Hedgehog signal transduction functions at the mechanistic level using the mouse as a model system. Specifically, the roles of 2 novel antagonists of the mammalian pathway, Wdr10 and Tulp3, will be studied using genetic, cellular, and biochemical approaches outlined in 3 specific aims. The Wdr10 protein, as well as its invertebrate homologs, functions in intraflagellar transport, a process used for building cilia and flagella. The experiments proposed in Aim 1 will confirm the requirement for Wdr10 in regulating the Hedgehog pathway through targeted mutagenesis and transgenic rescue approaches. Other experiments will address the subcellular localization of Wdr10 and test whether it acts in retrograde transport using immunocytochemistry and live cell imaging techniques. The mechanism of Wdr10 function in the Hedgehog pathway will be addressed in Aim 2. The hypothesis that Wdr10 acts cell autonomously will be tested through chimera analysis and the step in the Hedgehog pathway at which Wdr10 acts will be clarified through epistasis analysis. In addition, the hypothesis that Wdr10 acts directly or indirectly in controlling nuclear localization of the Gli transcription factors will be pursued using immunohistochemical and biochemical methods. Finally, the experiments in Aim 3 will test 2 hypotheses regarding the function of Tulp3 in Hedgehog signaling. First, the importance of the interaction between Tulp3 and myosin Vb will be functionally tested by blocking myosin Vb activity in vivo. Second, the significance of regulated Tulp3 nuclear translocation in Hedgehog signaling will be examined in tissue culture cells
Keywords: ATPase, Actin-Activated; Address; Adenosine Triphosphatase, Myosin; Alleles; Allelomorphs; Animal Model; Animal Models and Related Studies; Biochemical; Biological Models; Birth Defects; Blastocytes; Blastomeres; Blood Coagulation Factor IV; Ca++ element; Calcium; Calcium-Dependent Activator Protein; Calcium-Dependent Regulator; Calmodulin; Cancers; Carrier Proteins; Cell Communication and Signaling; Cell Fate Control; Cell Fate Regulation; Cell Nucleus; Cell Signaling; Cell membrane; Cells; Chimera; Chimera organism; Cilia; Coagulation Factor IV; Congenital Abnormality; Congenital Anatomic Abnormality; Congenital Anatomical Abnormality; Congenital Defects; Congenital Deformity; Congenital Malformation; Cytoplasmic Membrane; Data; Defect; EC 2.7; Embryonic Cell; Epistasis; Epistasis, Genetic; Epistatic Deviation; Erinaceidae; Factor IV; Family member; Flagella; G Protein-Coupled Receptor Signaling; GPCR Signaling; GeneHomolog; Genetic; Genetic Alteration; Genetic Change; Genetic Epistasis; Genetic defect; Genetics-Mutagenesis; Gx Protein; Hedgehog (Hh) signal transduction pathway; Hedgehogs; Homolog; Homologous Gene; Homologue; Human; Human, General; Imaging Procedures; Imaging Techniques; Interaction Deviation; Intracellular Communication and Signaling; Intracellular Transport; Invertebrata; Invertebrates; Invertebrates, General; Investigators; KRP protein; KRP(130); Kinases; Life; Ligands; Malignant Neoplasms; Malignant Tumor; Mammalia; Mammals; Mammals, General; Mammals, Mice; Man (Taxonomy); Man, Modern; Mediating; Methods; Mice; Model System; Models, Biologic; Molecular Biology, Mutagenesis; Molecular Genetic Abnormality; Motor; Murine; Mus; Mutagenesis; Mutation; Myosin ATPase; Myosin Adenosinetriphosphatase; Myosin Type V; Myosin V; Myosins; Nuclear; Nuclear Translocation; Nucleus; Pathway interactions; Phosphodiesterase Activating Factor; Phosphodiesterase Protein Activator; Phosphotransferases; Plasma Membrane; Play; Process; Programs (PT); Programs [Publication Type]; Proteins; Research; Research Personnel; Researchers; Role; SMO; SMO protein, human; SMOH; SMOH protein, human; Signal Transduction; Signal Transduction Systems; Signaling; Smoothened; Smoothened Homolog; SpKRP 85; SpKRP 95; Structure of blastomere; Technics, Imaging; Testing; Transgenic Organisms; Transphosphorylases; Transport Proteins; Transporter Protein; biological signal transduction; blastomere structure; cell imaging; cellular imaging; experiment; experimental research; experimental study; gene product; gene x gene interaction; genetic epistases; genome mutation; hedgehog signal transduction; hedgehog signaling pathway; hh signal transduction; hh signaling pathway; human SMO protein; immunocytochemistry; in vivo; insight; kinase-related protein; kinesin II; malignancy; model organism; mutant; myosin ATP phosphohydrolase (actin translocating); neoplasm/cancer; novel; pathway; plasmalemma; programs; research study; response; retrograde transport; smoothened homolog (Drosophila), human; smoothened signaling pathway; social role; tissue/cell culture; trafficking; transcription factor; transgenic
Project start date: 2005-07-15
Project end date: 2010-06-30
Budget start date: 1-JUL-2009
Budget end date: 30-JUN-2010
5R01HD050761-05 (2009): $289737
5R01HD050761-03 (2007): $287026
5R01HD050761-02 (2006): $295599
Grants awarded to Jonathan T Eggenschwiler
HEDGEHOG SIGNALING AND INTRACELLULAR TRANSPORT
Jonathan T Eggenschwiler, Assistant Professor
Princeton University, Office Of Research And Project Administration, Princeton, Nj 08544-0036
Grant 3R01HD050761-05S1 from Eunice Kennedy Shriver National Institute Of Child Health & Human Development
Abstract: Embryonic cells use the Hedgehog signaling pathway to convey information that controls cell fate, proliferation, and survival. Misregulation of this pathway leads to birth defects and a variety of cancers in humans. Recent data point to key mechanisms controlling the Hedgehog pathway in mammals that do not appear to be evolutionary conserved in invertebrate model organisms. The primary objective of the proposed research is to understand how Hedgehog signal transduction functions at the mechanistic level using the mouse as a model system. Specifically, the roles of 2 novel antagonists of the mammalian pathway, Wdr10 and Tulp3, will be studied using genetic, cellular, and biochemical approaches outlined in 3 specific aims. The Wdr10 protein, as well as its invertebrate homologs, functions in intraflagellar transport, a process used for building cilia and flagella. The experiments proposed in Aim 1 will confirm the requirement for Wdr10 in regulating the Hedgehog pathway through targeted mutagenesis and transgenic rescue approaches. Other experiments will address the subcellular localization of Wdr10 and test whether it acts in retrograde transport using immunocytochemistry and live cell imaging techniques. The mechanism of Wdr10 function in the Hedgehog pathway will be addressed in Aim 2. The hypothesis that Wdr10 acts cell autonomously will be tested through chimera analysis and the step in the Hedgehog pathway at which Wdr10 acts will be clarified through epistasis analysis. In addition, the hypothesis that Wdr10 acts directly or indirectly in controlling nuclear localization of the Gli transcription factors will be pursued using immunohistochemical and biochemical methods. Finally, the experiments in Aim 3 will test 2 hypotheses regarding the function of Tulp3 in Hedgehog signaling. First, the importance of the interaction between Tulp3 and myosin Vb will be functionally tested by blocking myosin Vb activity in vivo. Second, the significance of regulated Tulp3 nuclear translocation in Hedgehog signaling will be examined in tissue culture cells
Keywords: ATPase, Actin-Activated; Address; Adenosine Triphosphatase, Myosin; Alleles; Allelomorphs; Animal Model; Animal Models and Related Studies; Biochemical; Biological Models; Birth Defects; Blastocytes; Blastomeres; Blood Coagulation Factor IV; Ca++ element; Calcium; Calcium-Dependent Activator Protein; Calcium-Dependent Regulator; Calmodulin; Cancers; Carrier Proteins; Cell Communication and Signaling; Cell Fate Control; Cell Fate Regulation; Cell Nucleus; Cell Signaling; Cell membrane; Cells; Chimera; Chimera organism; Cilia; Coagulation Factor IV; Congenital Abnormality; Congenital Anatomic Abnormality; Congenital Anatomical Abnormality; Congenital Defects; Congenital Deformity; Congenital Malformation; Cytoplasmic Membrane; Data; Defect; EC 2.7; Embryonic Cell; Epistasis; Epistasis, Genetic; Epistatic Deviation; Erinaceidae; Factor IV; Family member; Flagella; G Protein-Coupled Receptor Signaling; GPCR Signaling; GeneHomolog; Genetic; Genetic Alteration; Genetic Change; Genetic Epistasis; Genetic defect; Genetics-Mutagenesis; Gx Protein; Hedgehog (Hh) signal transduction pathway; Hedgehogs; Homolog; Homologous Gene; Homologue; Human; Human, General; Imaging Procedures; Imaging Techniques; Interaction Deviation; Intracellular Communication and Signaling; Intracellular Transport; Invertebrata; Invertebrates; Invertebrates, General; Investigators; KRP protein; KRP(130); Kinases; Life; Ligands; Malignant Neoplasms; Malignant Tumor; Mammalia; Mammals; Mammals, General; Mammals, Mice; Man (Taxonomy); Man, Modern; Mediating; Methods; Mice; Model System; Models, Biologic; Molecular Biology, Mutagenesis; Molecular Genetic Abnormality; Motor; Murine; Mus; Mutagenesis; Mutation; Myosin ATPase; Myosin Adenosinetriphosphatase; Myosin Type V; Myosin V; Myosins; Nuclear; Nuclear Translocation; Nucleus; Pathway interactions; Phosphodiesterase Activating Factor; Phosphodiesterase Protein Activator; Phosphotransferases; Plasma Membrane; Play; Process; Programs (PT); Programs [Publication Type]; Proteins; Research; Research Personnel; Researchers; Role; SMO; SMO protein, human; SMOH; SMOH protein, human; Signal Transduction; Signal Transduction Systems; Signaling; Smoothened; Smoothened Homolog; SpKRP 85; SpKRP 95; Structure of blastomere; Technics, Imaging; Testing; Transgenic Organisms; Transphosphorylases; Transport Proteins; Transporter Protein; biological signal transduction; blastomere structure; cell imaging; cellular imaging; experiment; experimental research; experimental study; gene product; gene x gene interaction; genetic epistases; genome mutation; hedgehog signal transduction; hedgehog signaling pathway; hh signal transduction; hh signaling pathway; human SMO protein; immunocytochemistry; in vivo; insight; kinase-related protein; kinesin II; malignancy; model organism; mutant; myosin ATP phosphohydrolase (actin translocating); neoplasm/cancer; novel; pathway; plasmalemma; programs; research study; response; retrograde transport; smoothened homolog (Drosophila), human; smoothened signaling pathway; social role; tissue/cell culture; trafficking; transcription factor; transgenic
Project start date: 2009-09-30
Project end date: 2010-06-29
Budget start date: 30-SEP-2009
Budget end date: 29-JUN-2010
3R01HD050761-05S1 (2009): $66152
1R01HD050761-01 (2005): $324000