MECHANISMS OF STEROID-TRIGGERED PROGRAMMED CELL DEATH
Carl S Thummel, Investigator
University Of Utah, 75 South 2000 East, Salt Lake City, Ut 84112
Grant 3R01GM073670-04S1 from National Institute Of General Medical Sciences
Abstract: Small lipophilic hormones, acting through their corresponding nuclear receptors, control a wide range of developmental and physiological responses in higher organisms. Although extensive studies have focused on the mechanisms by which nuclear receptors control target gene transcription, relatively little is known about how a hormonal signal is transduced into an appropriate biological response during development. We propose to define one such pathway in detail - steroid-triggered cell death - using Drosophila as a model system. It is well known that steroids play a central role in controlling cell death in higher organisms, including humans. Only in Drosophila, however, has a genetic cascade been identified that links the hormone to a death response - the destruction of the larval salivary glands in response to the steroid hormone ecdysone during metamorphosis. We propose to build off this foundation, using an open-ended genetic screen to identify key players in this pathway. By using GFP as a marker for salivary glands in living animals, we will identify mutants that show specific defects in the steroid-triggered death response. A pilot screen has demonstrated the feasibility of this approach. We identified known genes in the death pathway as well as several new players, including genes that encode the CBP transcriptional co-factor and the TBP-related factor, TRF2. We propose to characterize these two genes in detail, defining the mechanisms that link them to cell death. We also propose to expand our search for death regulators through saturation mutagenesis of approximately 40% of the genome. Mutations will be mapped to specific genes, and functions for these genes will be assigned. This work provides a basis for understanding the molecular mechanisms of hormone signal transduction - defining the players in a genetic cascade that link the hormone to a stage- and tissue-specific biological response during development. This work also represents the first attempt to use random mutagenesis to dissect an endogenous programmed cell death response in Drosophila, raising the possibility that we will uncover novel death regulators. Finally, our studies provide a foundation for determining how steroids control cell death in humans, with implications for understanding and treating human disease
Keywords: Affect; Alleles; Allelomorphs; Animals; Apoptosis; Apoptosis Pathway; Biological; Biological Metamorphosis; Biological Models; Body Tissues; Cell Communication and Signaling; Cell Death; Cell Death, Programmed; Cell Signaling; Cells; Cessation of life; Chemotherapy-Hormones/Steroids; Cholest-7-en-6-one, 2, 3, 14, 22, 25-pentahydroxy-, (2beta, 3beta, 5beta, 22R)-; Chromosomes; DIAP1 protein, Drosophila; Death; Defect; Development; Disease; Disorder; Down-Regulation; Down-Regulation (Physiology); Downregulation; Drosophila; Drosophila genus; Drosophila inhibitor of apoptosis 1; EP300; EP300 gene; Ecdysone; Endocrine Gland Secretion; Event; Foundations; Fruit Fly, Drosophila; Gene Transcription; Genes; Genetic; Genetic Alteration; Genetic Change; Genetic Screening; Genetic Transcription; Genetic defect; Genetics-Mutagenesis; Genome; Goals; Head and Neck, Salivary Glands; Hormonal; Hormones; Human; Human, General; IAP1, Drosophila; Intracellular Communication and Signaling; Investigators; Lead; Life; Link; Man (Taxonomy); Man, Modern; Maps; Metamorphosis, Biological; Model System; Models, Biologic; Molecular; Molecular Biology, Mutagenesis; Molting Hormone; Mutagenesis; Mutation; Nuclear Receptors; Organism; Ortholog; Orthologous Gene; Pathway interactions; Pb element; Phenotype; Physiologic; Physiological; Play; Programs (PT); Programs [Publication Type]; RNA Expression; Regulation; Research Personnel; Researchers; Role; Salivary Glands; Signal Transduction; Signal Transduction Systems; Signaling; Sorting - Cell Movement; Staging; Steroid Compound; Steroids; TATA Box Binding Protein-Like Proteins; TATA-Binding Protein-Related Factors; TBP-Like Protein; TBP-Like TLP; TBP-Related Factor; TLP; Testing; Therapeutic Hormone; Therapeutic Steroid Hormone; Tissues; Transcription; Transcription, Genetic; Work; base; biological signal transduction; disease/disorder; fruit fly; gene function; genome mutation; heavy metal Pb; heavy metal lead; human disease; inhibitor of apoptosis 1, Drosophila; living system; metamorphosis; mutant; necrocytosis; novel; p300; pathway; programs; response; social role; sorting; steroid hormone; tool; transcription factor
Project start date: 2009-08-13
Project end date: 2010-07-31
Budget start date: 13-AUG-2009
Budget end date: 31-JUL-2010
3R01GM073670-04S1 (2009): $14282
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Carl S Thummel
DEVELOPMENTAL BIOLOGY TRAINING PROGRAM
Carl S Thummel, Professor
University Of Utah 75 South 2000 East Salt Lake City, Ut 84112
Grant 5T32HD007491-10 from National Institute Of Child Health And Human Development IRG: CHHD
Abstract: This application requests continued support for an interdepartmental Developmental Biology Training Program at the University of Utah. This Program provides support for training exceptional predoctoral and postdoctoral scientists in the field of Developmental Biology. The Program consists of individualized research training under the guidance of 46 faculty members who work in one of five Ph.D. degree-granting departments within the University. The Program Director and interdepartmental Advisory Committee members select Trainees, monitor their progress, and organize Training Program activities. Training is provided in a broad range of areas including gene regulation, cell differentiation, growth and morphogenesis, signal transduction, and developmental genetics. Prospective Ph.D. Trainees are admitted to graduate school through the Molecular Biology and Neuroscience Graduate Programs, which have interdepartmental admission committees. During their first two years, Ph.D. students take a series of interdepartmental core courses, perform laboratory rotations, act as teaching assistants, and Complete their Ph.D. qualifying exams. A research advisor and thesis project are chosen at the end of the first year, and all trainees must satisfy the particular Ph.D. degree requirements of their home department. Predoctoral and Postdoctoral trainees are selected based on excellence in research and are supported for up to three years for predoctoral trainees and two years for postdoctoral trainees. The participation of five departments provides a diverse interdisciplinary training in Developmental Biology. A coherent structure is provided by the wide variety of interdepartmental activities fostered by the Molecular Biology and Neurosciences Programs as well as the long history of cooperation and collaboration within the University community. All trainees are required to take a Scientific Ethics course, participate in an ongoing Journal Club related to developmental biology, take an advanced course in developmental biology, participate in and present a seminar in the Developmental Biology Discussion Group, participate in and present a research- based talk at an annual Training Program retreat, and host an outside seminar speaker. This is supplemented with vigorous seminar programs and inter- laboratory research-in-progress group meetings to ensure that our trainees receive a strong training in developmental biology, preparing them to direct their own first-rate independent research programs.
Project start date: 1995-09-29
Project end date: 2006-04-30
5T32HD007491-10 (2005): $313075
5T32HD007491-09 (2004): $244336
5T32HD007491-08 (2003): $288221
5T32HD007491-07 (2002): $295151
2T32HD007491-06 (2001): $264254
REGULATION AND FUNCTION OF DROSOPHILA NUCLEAR RECEPTORS
Carl S Thummel, Investigator
University Of Utah, 75 South 2000 East, Salt Lake City, Ut 84112
Grant 3R01DK075607-04S1 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: This application is in response to Notice Number NOT-OD-09-058 NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications, and represents an expansion of our currently funded grant 5R01 DK075607-03; Regulation and Function of Drosophila Nuclear Receptors. The specific aims of the parent grant addressed two major nuclear receptor-regulated pathways adult maturation and metabolism. We have completed our studies of maturation and are now focusing our attention on roles for Drosophila nuclear receptors (NRs) in sensing metabolites and maintaining metabolic homeostasis, with the goal of identifying and characterizing the critical aspects of NR signaling that are conserved through evolution up to humans. In this Competitive Revision application, we propose to accelerate our studies by characterizing the metabolic functions of two Drosophila NRs DHR3, the single fly ortholog of the mammalian ROR subclass, and DHR38, the fly ortholog of mammalian NR4A receptors. Our preliminary data indicate that each of these receptors has a critical role in metabolism that is similar to that of its vertebrate counterpart. DHR3 binds cholesterol and its activation depends on sterol levels in vivo. In addition, several key sterol metabolic genes are misregulated in DHR3 mutants, and DHR3 mutants have reduced cholesterol levels. These observations lead us to propose that DHR3 functions as a cholesterol sensor to maintain cholesterol homeostasis. For DHR38, null mutants have normal levels of circulating sugar and triacylglycerol, but display reduced levels of glycogen and has reduced expression of phosphoglucomutase, a key enzyme that acts in glycogenolysis and glycogenesis. These observations support the hypothesis that DHR38 plays a critical role in maintaining carbohydrate homeostasis. To test these hypotheses, we will conduct detailed phenotypic and metabolic characterization of DHR3 and DHR38 mutants. We will study the roles of signaling pathways and ligands in controlling NR activity. Finally, we will identify and characterize target genes that are directly regulated by each NR and that contribute to key metabolic activities of the receptor. These experiments follow along the lines of study proposed in the parent grant - using Drosophila as a model system to define the molecular mechanisms of NR regulation and function, with direct implications for how the orthologous NRs act in humans, as well as their contributions to critical human diseases associated with NR dysfunction, including cardiovascular disease, diabetes, and obesity. Our studies use Drosophila as a simple model system to define the molecular mechanisms of nuclear receptor action that are conserved through evolution up to humans. This work will have an impact on our understanding of normal nuclear receptor signaling pathways and provide new directions for combating critical human diseases associated with nuclear receptor dysfunction, including cardiovascular disease, diabetes, and obesity
Keywords: 20-Hydroxyecdysone; 21+ years old; 3`5`-cyclic ester of AMP; 9-cis-Retinoic Acid Receptor; Address; Adenosine Cyclic 3`, 5`-Monophosphate; Adenosine Cyclic Monophosphate; Adenosine, cyclic 3`, 5`-(hydrogen phosphate); Adipocytes; Adipose Cell; Adult; Animals; Apoptosis; Apoptosis Pathway; Area; Atheroscleroses; Atherosclerosis; Atherosclerotic Cardiovascular Disease; Attention; Autoregulation; Beta-Ecdysone; Binding; Binding (Molecular Function); Biological Models; Blood Serum; Body Tissues; Carbohydrates; Cardiovascular Diseases; Cell Communication and Signaling; Cell Death, Programmed; Cell Signaling; Cholest-5-en-3-ol (3beta)-; Cholest-7-en-6-one, 2, 3, 14, 20, 22, 25-hexahydroxy-, (2beta, 3beta, 5beta, 22R)-; Cholesterol; Cholesterol Homeostasis; Collection; Crustecdysone; Cyclic AMP; D-Glucose; Data; Dextrose; Diabetes Mellitus; Drosophila; Drosophila genus; Dysfunction; Ecdysterone; Embryo; Embryonic; Enzymes; Evolution; Family member; Fat Cells; Flies; Fruit Fly, Drosophila; Functional disorder; Funding; Gene Targeting; Genes; Genetic; Genomics; Gluconeogenesis; Glucose; Glucose Phosphomutase; Glycogen; Goals; Grant; Hepatic Cells; Hepatic Parenchymal Cell; Hepatocyte; Homeostasis; Human; Human, Adult; Human, General; Intermediary Metabolism; Intracellular Communication and Signaling; Lead; Ligands; Link; Lipocytes; Liver Cells; METBL; Mammals, Mice; Man (Taxonomy); Man, Modern; Mature Lipocyte; Mature fat cell; Metabolic; Metabolic Processes; Metabolism; Metabolism, Carbohydrates/Storage/Polysaccharides; Mice; Model System; Models, Biologic; Molecular; Molecular Interaction; Murine; Mus; NIH; National Institutes of Health; National Institutes of Health (U.S.); Nuclear Receptors; Obesity; Ortholog; Orthologous Gene; Pathway interactions; Pb element; Phosphoglucomutase; Physiological Homeostasis; Physiopathology; Play; Population; Predisposition; Programs (PT); Programs [Publication Type]; RXR; RXR Protein; Receptor Protein; Receptor Signaling; Recovery; Regulation; Retinoic Acid Receptor RXR; Retinoid X Receptors; Role; Serum; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Sterols; Susceptibility; Targetings, Gene; Testing; Therapeutic Steroid Hormone; Tissues; Trehalose; Triacylglycerol; Triglycerides; United States National Institutes of Health; Work; adenosine 3`5` monophosphate; adiposity; adult human (21+); alpha-D-Glucopyranoside, alpha-D-glucopyranosyl; alpha-D-Glucose 1, 6-phosphomutase; atheromatosis; atherosclerotic vascular disease; biological signal transduction; cAMP; carbohydrate metabolism; cardiovascular disorder; cholesterol control; cholesterol management; cholesterol metabolism; combat; corpulence; corpulency; corpulentia; diabetes; enzyme activity; experiment; experimental research; experimental study; fly; fruit fly; glucose biosynthesis; glycogenesis; glycogenolysis; heavy metal Pb; heavy metal lead; human disease; in vivo; insulin sensitivity; manage cholesterol; member; mutant; neuron development; obese; obese people; obese person; obese population; parent grant; pathophysiology; pathway; programs; public health relevance; receptor; research study; response; sensor; social role; steroid hormone; sugar; tool
Relevance: Our studies use Drosophila as a simple model system to define the molecular mechanisms of nuclear receptor action that are conserved through evolution up to humans. This work will have an impact on our understanding of normal nuclear receptor signaling pathways and provide new directions for combating critical human diseases associated with nuclear receptor dysfunction, including cardiovascular disease, diabetes, and obesity
Project start date: 2006-07-01
Project end date: 2011-05-31
Budget start date: 12-SEP-2009
Budget end date: 31-MAY-2011
PFA/PA: PA-07-070
3R01DK075607-04S1 (2009): $430823
A DROSOPHILA MODEL FOR GENETIC STUDIES OF METABOLISM
Carl S Thummel
University Of Utah, 75 South 2000 East, Salt Lake City, Ut 84112
Grant 5RC1DK086426-02 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: This application addresses the broad Challenge Area (15) Translational Science and the specific Challenge Topic 15-DK-102 Develop improved animal models of NIDDK diseases. Metabolic homeostasis plays a central role in all aspects of postembryonic life, allowing animals to balance their dietary intake with the energy needs required for day-to-day survival. Conversely, misregulation of metabolism can lead to obesity and type 2 diabetes, which are critical risk factors for human disease, including cardiovascular disorders and cancer. The goal of our research is to use the fruit fly, Drosophila melanogaster, as a simple model system to define the central regulatory pathways that control metabolism and maintain energy homeostasis in all higher organisms, including humans. Our experimental approach exploits the unique strengths of Drosophila as a model system by conducting an open-ended genetic screen in the intact animal. This proposal arises from our ongoing studies of nuclear receptor signaling in Drosophila and the central role of these factors in maintaining metabolic homeostasis. Genetic studies have demonstrated that the nuclear receptor DHR96 is required to maintain appropriate triacylglycerol (TAG) levels in the animal. DHR96 mutants are viable, have low levels of TAG, and are sensitive to starvation, correlating with the misregulation of key lipid metabolic genes. We have discovered that the starvation sensitivity of DHR96 mutants can be rescued by introducing second-site mutations in genes that increase TAG levels, such as the adipose triglyceride lipase gene or the adipokinetic hormone receptor gene (which acts like glucagon to drive lipolysis). This observation provides a framework for identifying new genes that control lipid metabolism. We propose to exploit the DHR96 mutant as a sensitized genetic background for conducting open-ended genetic screens with the aim of identifying mutations that rescue its starvation sensitivity. Our preliminary data indicate that this screen should uncover a range of genes involved in many aspects of lipid metabolism, including genes which, when mutated, lead to obesity. Recently developed methods for efficient single-gene disruption by transposon mutagenesis will facilitate the screen and allow rapid gene identification. These studies provide a new basis for using Drosophila as means of extending our understanding of key lipid metabolic pathways that impact human health. Genetic screens represent one of the most powerful and important advantages of working in Drosophila, and offer a way to expand our understanding of specific biological pathways in new and unexpected directions. This approach has had a major impact on our understanding of human health through the delineation of the fundamental pathways that dictate embryonic development and the discovery of central signaling pathways, such as Notch, Wnt, and hedgehog signaling. We propose to exploit this strength of the fly toward the discovery of novel genes that impact lipid metabolism. We will focus our studies on newly identified Drosophila genes that have close homologs in mice and humans to facilitate the translation of our discoveries into vertebrate systems. Our long-term goal is to provide new candidates for mouse gene knockout studies and human disease gene mapping. In this way we hope to use the fly as a tool for gene discovery and extend these advances toward a better understanding of the causes of human metabolic disorders. The dramatic rise in metabolic disorders, such as diabetes and obesity, poses a major health risk to the world population. This proposed research will use the fruit fly, Drosophila, as a simple genetic model system to define the fundamental mechanisms that control lipid metabolism, with the goal of providing new directions for understanding and treating human metabolic disorders
Keywords: Abnormal Assessment of Metabolism; Address; Adipose tissue; Affect; Animal Model; Animal Models and Related Studies; Animals; Antidiabetic Hormone; Area; Autoregulation; Biological; Biological Models; Cancers; Cardiovascular Diseases; Chromosome Mapping; Data; Diabetes Mellitus; Diabetes Mellitus, Adult-Onset; Diabetes Mellitus, Ketosis-Resistant; Diabetes Mellitus, Non-Insulin-Dependent; Diabetes Mellitus, Noninsulin Dependent; Diabetes Mellitus, Slow-Onset; Diabetes Mellitus, Stable; Diabetes Mellitus, Type 2; Diabetes Mellitus, Type II; Dietary intake; Disease; Disorder; Drosophila; Drosophila genus; Drosophila melanogaster; Embryo Development; Embryogenesis; Embryonic Development; Equilibrium; Fatty Tissue; Flies; Fruit Fly, Drosophila; GCG; Gene Localization; Gene Mapping; Gene Mapping, Total Human and Non-Human; GeneHomolog; Genes; Genetic; Genetic Alteration; Genetic Change; Genetic Models; Genetic Screening; Genetic defect; Genetics, Gene Mapping; Genetics-Mutagenesis; Glucagon; Glucagon (1-29); Glukagon; Goals; HG-Factor; Health; Hedgehog (Hh) signal transduction pathway; Homeostasis; Homolog; Homologous Gene; Homologue; Hormone Receptor; Human; Human, General; Hyperglycemic-Glycogenolytic Factor; Intermediary Metabolism; Lead; Life; Linkage Mapping; Lipase; Lipids; Lipolysis; METBL; MODY; Malignant Neoplasms; Malignant Tumor; Mammals, Mice; Man (Taxonomy); Man, Modern; Maps; Maturity-Onset Diabetes Mellitus; Metabolic; Metabolic Diseases; Metabolic Disorder; Metabolic Pathway; Metabolic Processes; Metabolic Studies; Metabolism; Metabolism Studies; Metabolism, Lipids/Lipoproteins/Membrane Constituents; Methods; Mice; Model System; Models, Biologic; Molecular Biology, Mutagenesis; Molecular Genetic; Molecular Genetics; Murine; Mus; Mutagenesis; Mutate; Mutation; NIDDK; NIDDM; National Institute of Diabetes and Digestive and Kidney Diseases; National Institute of Digestive Diseases and Kidney Disorders; Non-Insulin Dependent Diabetes; Non-Insulin-Dependent Diabetes Mellitus; Nuclear Receptors; Obesity; Organism; Pathway interactions; Pb element; Physiological Homeostasis; Play; Population; Receptor Gene; Receptor Signaling; Regulatory Pathway; Research; Risk; Risk Factors; Role; Screening procedure; Signal Pathway; Site; Starvation; System; System, LOINC Axis 4; T2D; T2DM; Thesaurismosis; Translational Research; Translational Research Enterprise; Translational Science; Translations; Triacylglycerol; Triacylglycerol Hydrolase; Triacylglycerol Lipase; Triacylglycerol acylhydrolase; Tributyrinase; Triglyceridase; Triglyceride Lipase; Triglycerides; Triolean Hydrolase; Type 2 diabetes; Type II diabetes; Work; adipokinetic hormone; adipose; adiposity; adult onset diabetes; balance; balance function; base; cardiovascular disorder; corpulence; corpulency; corpulentia; diabetes; disease/disorder; fat metabolism; fly; fruit fly; gene discovery; genetic mapping; genome mutation; heavy metal Pb; heavy metal lead; hedgehog signaling pathway; hh signaling pathway; human disease; improved; ketosis resistant diabetes; knockout gene; lipid metabolism; living system; malignancy; maturity onset diabetes; metabolic abnormality assessment; metabolism disorder; model organism; mutant; neoplasm/cancer; notch; notch protein; notch receptors; novel; obese; obese people; obese person; obese population; pathway; response; screening; screenings; smoothened signaling pathway; social role; tool; translation research enterprise; tributyrase; white adipose tissue; yellow adipose tissue
Relevance: The dramatic rise in metabolic disorders, such as diabetes and obesity, poses a major health risk to the world population. This proposed research will use the fruit fly, Drosophila, as a simple genetic model system to define the fundamental mechanisms that control lipid metabolism, with the goal of providing new directions for understanding and treating human metabolic disorders
Project start date: 2009-09-19
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: RFA-OD-09-003
5RC1DK086426-02 (2010): $389113
1RC1DK086426-01 (2009): $346269
REGULATION AND FUNCTION OF DROSOPHILA NUCLEAR RECEPTORS
Carl S Thummel, Investigator
University Of Utah, 75 South 2000 East, Salt Lake City, Ut 84112
Grant 3R01DK075607-04S2 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: The fruit fly Drosophila melanogaster has 18 nuclear receptor (NR) genes, significantly fewer than the 48 genes found in humans, spanning all vertebrate NR subclasses and encoding orthologs of key human receptors, including RXR (USP in flies), NGFI-B/NURR (DHR38), ERR (dERR), SXR (DHR96), and HNF4 (dHNF4). These characteristics of Drosophila NRs, combined with an extensive collection of genetic and genomic tools, establish the fly as an ideal model system for studying the molecular mechanisms of NR regulation and function. In this proposal, we focus on 2 major NR-regulated biological pathways steroid-triggered maturation and lipid metabolism. Like vertebrates, Drosophila maturation is triggered by steroid hormones and their receptors, via complex transcriptional cascades that were identified and characterized in the fly. In contrast, no studies to date have addressed roles for Drosophila NRs in lipid metabolism. We will study the steroid regulation of a poorly understood transition that occurs during the last larval stage, when the animal commits to terminating its juvenile growth phase and initiating maturation via metamorphosis. We will identify genes regulated by alpha-ecdysone (E, the primary secreted steroid in Drosophila and the precursor to the active hormone 20-hydroxyecdysone, or 20E) and ask if these effects go through the DHR38 E receptor and its RXR partner, USP. We will determine if DHR38 and EcR function as partially redundant steroid receptors to initiate maturation. We will determine if dERR is transcriptionally controlled by 20E, examine its possible roles in initiating metamorphosis, and determine whether it controls lipid or sterol metabolism. We will follow up on the observation that DHR96 co-purifies with cholesterol and that DHR96 null mutants require cholesterol for their survival on minimal medium. Finally, we will characterize the expression and function of dHNF4 to define its roles in lipid metabolism and development. Metabolic profiling and microarrays will be used to gain a more complete understanding of the molecular mechanisms of dERR, DHR96, and dHNF4 function. Selected genes will be defined as direct regulatory targets. These studies will expand our understanding of NR signaling pathways with direct implications for how the orthologous NRs function in humans, as well as their contributions to critical human diseases associated with NR dysfunction, including cardiovascular disease, diabetes, and obesity
Keywords: 2, 6-Piperidinedione, 4-(2-(3, 5-dimethyl-2-oxocyclohexyl)-2-hydroxyethyl)-, (1S-(1alpha(S*), 3alpha, 5beta))-; 20-Hydroxyecdysone; 21+ years old; 9-cis-Retinoic Acid Receptor; Active Follow-up; Address; Adolescent; Adolescent Youth; Adult; Animals; Antibodies; Autoregulation; Beta-Ecdysone; Biological; Biological Metamorphosis; Biological Models; C elegans; C.elegans; Caenorhabditis elegans; Cardiovascular Diseases; Characteristics; Chemotherapy-Hormones/Steroids; Cholest-5-en-3-ol (3beta)-; Cholest-7-en-6-one, 2, 3, 14, 20, 22, 25-hexahydroxy-, (2beta, 3beta, 5beta, 22R)-; Cholest-7-en-6-one, 2, 3, 14, 22, 25-pentahydroxy-, (2beta, 3beta, 5beta, 22R)-; Cholesterol; Cicloheximide; Collection; Commit; Complex; Crustecdysone; Cycloheximide; Data; Development; Diabetes Mellitus; Drosophila; Drosophila genus; Drosophila melanogaster; Dysfunction; Ecdysone; Ecdysterone; Endocrine Gland Secretion; Flies; Fruit Fly, Drosophila; Functional disorder; Gene Targeting; Gene Transcription; Generalized Growth; Genes; Genetic; Genetic Transcription; Genomics; Growth; Homeostasis; Hormones; Human; Human, Adult; Human, General; Intermediary Metabolism; Investigators; Lipids; METBL; Man (Taxonomy); Man, Modern; Mediating; Metabolic; Metabolic Pathway; Metabolic Processes; Metabolism; Metabolism, Lipids/Lipoproteins/Membrane Constituents; Metamorphosis, Biological; Model System; Modeling; Models, Biologic; Molecular; Molting Hormone; Nuclear Receptor Gene; Nuclear Receptors; Obesity; Ortholog; Orthologous Gene; Pathway interactions; Pattern; Phase; Physiologic pulse; Physiological Homeostasis; Physiopathology; Programs (PT); Programs [Publication Type]; Pulse; RNA Expression; RXR; RXR Protein; Receptor Protein; Receptor Signaling; Receptors, Steroid; Regulation; Research Personnel; Researchers; Retinoic Acid Receptor RXR; Retinoid X Receptors; Role; SXR; SXR receptor; Signal Pathway; Staging; Steroid Compound; Steroid Receptors; Steroids; Sterols; Targetings, Gene; Testing; Therapeutic Hormone; Therapeutic Steroid Hormone; Tissue Growth; Transcription; Transcription, Genetic; Vertebrate Animals; Vertebrates; adiposity; adult human (21+); cardiovascular disorder; corpulence; corpulency; corpulentia; diabetes; ecdysteroid receptor; ecdysterone receptor; fat metabolism; fly; follow-up; fruit fly; gain of function; gain of function mutation; human disease; in vivo; juvenile; juvenile human; lipid metabolism; loss of function; member; metamorphosis; mutant; obese; obese people; obese person; obese population; ontogeny; pathophysiology; pathway; programs; receptor; receptor, 20-hydroxyecdysone; reproductive; response; social role; steroid X receptor; steroid and xenobiotic receptor; steroid hormone; steroid hormone receptor; steroid metabolism; tool; vertebrata
Project start date: 2010-01-15
Project end date: 2010-03-31
Budget start date: 15-JAN-2010
Budget end date: 31-MAR-2010
3R01DK075607-04S2 (2010): $44897
5R01DK075607-05 (2010): $343979
Sponsored Links Excellgen http://Excellgen.com
5R01DK075607-04 (2009): $337335
5R01DK075607-02 (2007): $324458
1R01DK075607-01 (2006): $324415
MECHANISMS OF STEROID-TRIGGERED PROGRAMMED CELL DEATH
Carl S Thummel, Investigator
University Of Utah, 75 South 2000 East, Salt Lake City, Ut 84112
Grant 5R01GM073670-04 from National Institute Of General Medical Sciences
Abstract: Small lipophilic hormones, acting through their corresponding nuclear receptors, control a wide range of developmental and physiological responses in higher organisms. Although extensive studies have focused on the mechanisms by which nuclear receptors control target gene transcription, relatively little is known about how a hormonal signal is transduced into an appropriate biological response during development. We propose to define one such pathway in detail - steroid-triggered cell death - using Drosophila as a model system. It is well known that steroids play a central role in controlling cell death in higher organisms, including humans. Only in Drosophila, however, has a genetic cascade been identified that links the hormone to a death response - the destruction of the larval salivary glands in response to the steroid hormone ecdysone during metamorphosis. We propose to build off this foundation, using an open-ended genetic screen to identify key players in this pathway. By using GFP as a marker for salivary glands in living animals, we will identify mutants that show specific defects in the steroid-triggered death response. A pilot screen has demonstrated the feasibility of this approach. We identified known genes in the death pathway as well as several new players, including genes that encode the CBP transcriptional co-factor and the TBP-related factor, TRF2. We propose to characterize these two genes in detail, defining the mechanisms that link them to cell death. We also propose to expand our search for death regulators through saturation mutagenesis of approximately 40% of the genome. Mutations will be mapped to specific genes, and functions for these genes will be assigned. This work provides a basis for understanding the molecular mechanisms of hormone signal transduction - defining the players in a genetic cascade that link the hormone to a stage- and tissue-specific biological response during development. This work also represents the first attempt to use random mutagenesis to dissect an endogenous programmed cell death response in Drosophila, raising the possibility that we will uncover novel death regulators. Finally, our studies provide a foundation for determining how steroids control cell death in humans, with implications for understanding and treating human disease
Keywords: Affect; Alleles; Allelomorphs; Animals; Apoptosis; Apoptosis Pathway; Biological; Biological Metamorphosis; Biological Models; Body Tissues; Cell Communication and Signaling; Cell Death; Cell Death, Programmed; Cell Signaling; Cells; Cessation of life; Chemotherapy-Hormones/Steroids; Cholest-7-en-6-one, 2, 3, 14, 22, 25-pentahydroxy-, (2beta, 3beta, 5beta, 22R)-; Chromosomes; DIAP1 protein, Drosophila; Death; Deep; Defect; Depth; Development; Disease; Disorder; Down-Regulation; Down-Regulation (Physiology); Downregulation; Drosophila; Drosophila genus; Drosophila inhibitor of apoptosis 1; EP300; EP300 gene; Ecdysone; Endocrine Gland Secretion; Event; Foundations; Fruit Fly, Drosophila; GFP; Gene Transcription; Genes; Genetic; Genetic Alteration; Genetic Change; Genetic Screening; Genetic Transcription; Genetic defect; Genetics-Mutagenesis; Genome; Goals; Green Fluorescent Proteins; Head and Neck, Salivary Glands; Hormonal; Hormones; Human; Human, General; IAP1, Drosophila; Intracellular Communication and Signaling; Investigators; Lead; Life; Link; Man (Taxonomy); Man, Modern; Maps; Metamorphosis, Biological; Model System; Models, Biologic; Molecular; Molecular Biology, Mutagenesis; Molting Hormone; Mutagenesis; Mutation; Nuclear Receptors; Organism; Ortholog; Orthologous Gene; Pathway interactions; Pb element; Personal Satisfaction; Phenotype; Physiologic; Physiological; Play; Programs (PT); Programs [Publication Type]; RNA Expression; Range; Regulation; Research Personnel; Researchers; Role; Salivary Glands; Signal Transduction; Signal Transduction Systems; Signaling; Sorting - Cell Movement; Staging; Steroid Compound; Steroids; TATA Box Binding Protein-Like Proteins; TATA-Binding Protein-Related Factors; TBP-Like Protein; TBP-Like TLP; TBP-Related Factor; TLP; Testing; Therapeutic Hormone; Therapeutic Steroid Hormone; Tissues; Transcription; Transcription, Genetic; Work; base; biological signal transduction; disease/disorder; fruit fly; gene function; genome mutation; heavy metal Pb; heavy metal lead; human disease; inhibitor of apoptosis 1, Drosophila; living system; metamorphosis; mutant; necrocytosis; novel; p300; pathway; programs; response; social role; sorting; steroid hormone; tool; transcription factor; well-being
Project start date: 2005-04-01
Project end date: 2010-08-31
Budget start date: 1-SEP-2008
Budget end date: 31-AUG-2010
5R01GM073670-04 (2008): $0
5R01GM073670-03 (2007): $354383
5R01GM073670-02 (2006): $273674
1R01GM073670-01 (2005): $171925
STEROID REGULATION OF PROGRAMMED CELL DEATH
Carl S Thummel, Professor
Institute Of Human Geneticsuniversity Of Utah
75 South 2000 East
salt Lake City, Ut 84112
Grant 5R01GM060954-02 from National Institute Of General Medical Sciences IRG: END
Abstract: adapted from the proposal) This proposal is aimed at defining the molecular mechanisms by which the steroid hormone ecdysone regulates programmed cell death in Drosophila. The larval midgut and salivary glands undergo massive stage-specific apoptosis in response to two sequential pulses of ecdysone during metamorphosis. Entry of these tissues into programmed cell death is foreshadowed by the induction of two key death inducer genes, reaper and hid. The goal of this proposal is to identify the ecdysone-inducible transcription factors that directly regulate stage-specific reaper and hid expression. It has been shown that the ecdysone-inducible transcription factors encoded by the Broad-Complex and E74 are required for salivary gland cell death as well as reaper and hid transcription. It will be determined whether this is a direct regulatory link. It will also be determined whether the Broad-Complex and E74 play a similar role in the earlier induction of reaper and hid in doomed larval midguts. The hypothesis that stage-specific Rel/NF-kB and AP-1 signaling converge with the steroid signal to determine the appropriate temporal regulation of cell death will be tested. Finally, the minimal sequences in the reaper and hid promoters that are required for proper hormone-induced stage-specific transcription will be identified and these sequences will be used to identify other factors that contribute to this regulation. This proposal provides a means of understanding the molecular mechanisms by which nuclear receptors, NF-kB and AP-1 converge to direct a programmed cell death response. In addition, this study will provide insight into the molecular mechanisms by which systemic hormonal signals are refined into stage- and tissue-specific biological responses during development
Keywords: ecdysone, gene induction /repression, hormone regulation /control mechanism, programmed cell death, transcription factor AP1 protein, developmental genetics, nuclear factor kappa beta, nuclear receptor, salivary gland Drosophilidae, larva
Project start date: 2000-03-01
Project end date: 2004-02-29
5R01GM060954-02 (2001): $149834
1R01GM060954-01 (2000): $112125
TRANSCRIPTIONAL CONTROL OF DROSOPHILA DETOXIFICATION GENES
Carl S Thummel, Investigator
University Of Utah, 75 South 2000 East, Salt Lake City, Ut 84112
Grant 5R01GM079197-03 from National Institute Of General Medical Sciences
Keywords: 2, 4, 6(1H, 3H, 5H)-Pyrimidinetrione, 5-ethyl-5-phenyl-; 5` Flanking Region; 5` Flanking Sequence; Affect; Animals; Autoregulation; Binding Sites; Bio-Informatics; Biochemical; Bioinformatics; Biological Function; Biological Models; Biological Process; Cancers; Chemicals; Combining Site; Complex; DNA Binding; DNA Binding Interaction; DNA-Binding Proteins; Development; Drosophila; Drosophila genus; Drosophila melanogaster; Drug Industry; Drug Metabolic Detoxication; Drug Transport; Drugs; Elements; Environment; Enzymes; Evolution; Excretory function; Flies; Foundations; Fruit Fly, Drosophila; Gene Action Regulation; Gene Expression; Gene Expression Regulation; Gene Regulation; Gene Regulation Process; Gene Transcription; Genes; Genetic; Genetic Models; Genetic Screening; Genetic Transcription; Goals; Heat Shock; Heat-Shock Reaction; Heat-Shock Response; Homeostasis; Human; Human, General; Hypoxia; Hypoxic; Industry, Pharmaceutic; Infection; Insecta; Insects; Invertebrates, Insects; Life; Lung diseases; Malignant Neoplasms; Malignant Tumor; Mammals, Mice; Man (Taxonomy); Man, Modern; Maps; Mediating; Medication; Metabolic Detoxication, Drug; Metabolic Detoxification, Drug; Metabolic Drug Detoxications; Metabolism of Toxic Agents; Mice; Model System; Models, Biologic; Models, Genetic; Molecular; Murine; Mus; Organism; Ortholog; Orthologous Gene; Oxidative Stress; Oxygen Deficiency; PXR receptor; Pesticides; Pharmaceutic Preparations; Pharmaceutical Agent; Pharmaceutical Industry; Pharmaceutical Preparations; Pharmaceuticals; Pharmacologic Substance; Pharmacological Substance; Phenemal; Phenobarbital; Phenobarbitone; Phenylbarbital; Phenylethylbarbituric Acid; Physiological Homeostasis; Plants; Plants, General; Poisons; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Pulmonary Diseases; Pulmonary Disorder; RNA Expression; Reactive Site; Regulation; Research; Resistance; Respiratory Disease; Respiratory Disorder; Respiratory System Disease; Respiratory System Disorder; Response Elements; Risk Factors; Role; Scanning; Specificity; Testing; Toxic Chemical; Toxic Substance; Toxin; Trans-Acting Factors; Trans-Activators; Transactivators; Transcription; Transcription Regulation; Transcription, Genetic; Transcriptional Control; Transcriptional Regulation; Transgenic Animals; Xenobiotics; combat; coping; defense response; detoxification; drug/agent; excretion; experiment; experimental research; experimental study; fly; fruit fly; insight; living system; lung disorder; malignancy; mutant; neoplasm/cancer; null mutation; poison; pollutant; pregnane X receptor; research study; resistant; response; sedative; social role; toxic compound; trans acting factor (genetic); transcription factor; yeast genetics
Project start date: 2008-05-10
Project end date: 2012-02-28
Budget start date: 1-MAR-2010
Budget end date: 28-FEB-2011
PFA/PA: PA-07-070
5R01GM079197-03 (2010): $134096
Sponsored Links Excellgen http://Excellgen.com
ECDYSONE REGULATED GENES IN DROSOPHILA
Carl S Thummel, Professor
Institute Of Human Geneticsuniversity Of Utah
75 South 2000 East
salt Lake City, Ut 84112
Grant 5R29GM040905-05 from National Institute Of General Medical Sciences IRG: GEN
Keywords: ecdysone, gene expression, hormone regulation /control mechanism, metamorphosis, molecular genetics DNA binding protein, cytogenetics, developmental genetics, genetic mapping, genetic transcription, steroid hormone Drosophilidae, complementary DNA, genetic library, in situ hybridization, molecular cloning, mutagen testing, nucleic acid hybridization, nucleic acid probe, nucleic acid sequence, organ culture, radionuclide
Project start date: 1988-12-01
Project end date: 1994-11-30
5R29GM040905-05 (1993): $129131
5R29GM040905-04 (1992): $124243