MECHANISM OF THE TRANSCRIPTIONAL RESPONSE TO TRANSITION METALS
Michael Thomas Marr, Assistant Professor
Brandeis University, 415 South St, Ms #116, Waltham, Ma 02454-9110
Grant 5R01GM085250-02 from National Institute Of General Medical Sciences
Abstract: Essential divalent transition metals such as zinc, copper, and iron play important structural and catalytic roles in over 300 proteins. However, these and other transition metals also pose a potential threat to an organism. Left unchecked these metals can catalyze the generation of free radicals that damage all types of biological molecules. Transition metals must be kept in a physiological window. Diseases like Wilson´s disease and Menke´s disease demonstrate that either too much or too little metal can lead to pathologies of the liver, kidney, nervous system and connective tissues. Clearly metal homeostasis is an important aspect of cellular function. A major part of this control occurs at the level of transcription. One of the central players in this regulation is the metal response element binding protein (MTF-1). MTF-1 is a sequence specific DNA binding protein that perceives the metal status of a cell and activates genes accordingly. A good deal is known about MTF-1 itself, but very little is known about the protein co-factors that help MTF-1 efficiently activate transcription. We will use an RNAi based screen, in the Drosophila model system, to determine what protein co-factors are required for metal stimulated transcription. In addition we will characterize the physical and functional interactions between MTF-1 and these protein co-factors using a combination of cell based and in vitro assays. Finally we will identify the MTF-1 binding sites across the Drosophila genome to define the MTF-1 regulon that responds to metal stimuli. The long-term objective of our studies is to understand how a cell, in response to a diverse set of metals, differentially regulates the appropriate metal responsive genes to control metal homeostasis. Humans have a nutritional requirement for essential metals for normal growth and health. But in high doses many of these metals become toxic. Understandably, there is also a need to keep these metals in balance. We are studying the genes that help maintain this equilibrium. The long term goal is to understand how deregulation of these same genes contributes to metal induced disease
Keywords: Address; Affect; Autoregulation; Binding; Binding (Molecular Function); Binding Proteins; Binding Sites; Biological; Biological Models; Brain; Cadmium; Cd element; Cd thionein; Cell Communication and Signaling; Cell Function; Cell Process; Cell Signaling; Cell physiology; Cells; Cellular Function; Cellular Physiology; Cellular Process; Cerebral Pseudosclerosis; Cognitive Discrimination; Combining Site; Complex; Connective Tissue; Copper; Cu element; Discrimination; Discrimination (Psychology); Disease; Disorder; Dose; Drosophila; Drosophila genome; Drosophila genus; Elongation Factor; Encephalon; Encephalons; Equilibrium; Fe element; Free Radicals; Fruit Fly, Drosophila; Gene Action Regulation; Gene Expression Profile; Gene Expression Regulation; Gene Regulation; Gene Regulation Process; Gene Targeting; Gene Transcription; Generalized Growth; Generations; Genes; Genetic Transcription; Genomics; Goals; Gowers` chorea; Growth; Health; Hepato-Neurologic Wilson Disease; Hepatocerebral Degeneration; Hepatolenticular Degeneration; Hg element; Homeostasis; Human; Human, General; Hypocupremia, Congenital; Hypocupremias, Congenital; Immune Precipitation; Immunoprecipitation; Individual; Intracellular Communication and Signaling; Iron; Kidney; Kinky Hair Disease; Kinky Hair Syndrome; Kinnier-Wilson Disease; Knowledge; Lead; Left; Libraries; Ligand Binding Protein; Liver; MRE-binding transcription factor; MTF-1 protein; Mammalian Cell; Man (Taxonomy); Man, Modern; Mediator; Mediator of Activation; Mediator of activation protein; Menkes Kinky Hair Syndrome; Menkes Syndrome; Menkes disease; Menkes kinky hair syndrome (MKHS); Menkes` Disease; Mercury; Metabolic; Metallothionein; Metals; Microarray Analysis; Microarray-Based Analysis; Model System; Modeling; Models, Biologic; Molecular Interaction; NRVS-SYS; Nature; Nervous System; Nervous System, Brain; Nervous system structure; Neurohepatic Degeneration; Neurologic Body System; Neurologic Organ System; Normal Cell; Nutrient; Nutritional Requirements; Organism; Pathology; Pb element; Physiologic; Physiological; Physiological Homeostasis; Play; Position; Positioning Attribute; Post-Transcriptional Gene Silencing; Post-Transcriptional Gene Silencings; Posttranscriptional Gene Silencing; Posttranscriptional Gene Silencings; Programs (PT); Programs [Publication Type]; Progressive Lenticular Degeneration; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Proteins; Pseudosclerosis; Quelling; RNA Expression; RNA Interference; RNA Silencing; RNA Silencings; RNAi; Reactive Site; Regulation; Regulon; Response Elements; Role; Sequence-Specific DNA Binding Protein; Sequence-Specific Posttranscriptional Gene Silencing; Signal Transduction; Signal Transduction Systems; Signaling; Staging; Steely Hair Disease; Steely Hair Syndrome; Stimulus; Stress; Subcellular Process; System; System, LOINC Axis 4; Targetings, Gene; Tissue Growth; Toxic effect; Toxicities; Transcript; Transcription; Transcription Activation; Transcription Elongation; Transcription Initiation; Transcription Regulation; Transcription, Genetic; Transcriptional Activation; Transcriptional Control; Transcriptional Regulation; Transition Elements; Up-Regulation; Urinary System, Kidney; Westphal Strumpell disease; Westphal pseudosclerosis; Westphal-Strumpell Syndrome; Wilson Disease; Work; X-linked copper malabsorption; Zinc; Zn element; balance; balance function; base; biological signal transduction; body system, hepatic; cadmium metallothionein; cadmium-thionein; cohort; congenital hypocupraemia syndrome; copper transport disease; disease/disorder; familial hepatitis; fruit fly; gene expression signature; gene product; genome, Drosophila; genome, fruit fly; heavy metal Pb; heavy metal lead; in vitro Assay; interest; kinky hair syndrome (KHS); living system; metal-responsive transcription factor-1; microarray technology; nutrient requirement; ontogeny; organ system, hepatic; prevent; preventing; programs; public health relevance; renal; response; sensor; sex-linked neurodegenerative disease with monilethrix; social role; toxic metal; transcription factor; transcription factor MTF-1; transcriptome; transition metal; trichopoliodystrophy
Relevance: Humans have a nutritional requirement for essential metals for normal growth and health. But in high doses many of these metals become toxic. Understandably, there is also a need to keep these metals in balance. We are studying the genes that help maintain this equilibrium. The long term goal is to understand how deregulation of these same genes contributes to metal induced disease
Project start date: 2009-08-01
Project end date: 2014-05-31
Budget start date: 1-JUN-2010
Budget end date: 31-MAY-2011
PFA/PA: PA-07-070
5R01GM085250-02 (2010): $302527
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Michael Thomas Marr
MECHANISM OF THE TRANSCRIPTIONAL RESPONSE TO TRANSITION METALS
Michael Thomas Marr
Brandeis University, 415 South St, Ms #116, Waltham, Ma 02454-9110
Grant 3R01GM085250-02S1 from National Institute Of General Medical Sciences
Abstract: Essential divalent transition metals such as zinc, copper, and iron play important structural and catalytic roles in over 300 proteins. However, these and other transition metals also pose a potential threat to an organism. Left unchecked these metals can catalyze the generation of free radicals that damage all types of biological molecules. Transition metals must be kept in a physiological window. Diseases like Wilson´s disease and Menke´s disease demonstrate that either too much or too little metal can lead to pathologies of the liver, kidney, nervous system and connective tissues. Clearly metal homeostasis is an important aspect of cellular function. A major part of this control occurs at the level of transcription. One of the central players in this regulation is the metal response element binding protein (MTF-1). MTF-1 is a sequence specific DNA binding protein that perceives the metal status of a cell and activates genes accordingly. A good deal is known about MTF-1 itself, but very little is known about the protein co-factors that help MTF-1 efficiently activate transcription. We will use an RNAi based screen, in the Drosophila model system, to determine what protein co-factors are required for metal stimulated transcription. In addition we will characterize the physical and functional interactions between MTF-1 and these protein co-factors using a combination of cell based and in vitro assays. Finally we will identify the MTF-1 binding sites across the Drosophila genome to define the MTF-1 regulon that responds to metal stimuli. The long-term objective of our studies is to understand how a cell, in response to a diverse set of metals, differentially regulates the appropriate metal responsive genes to control metal homeostasis. Humans have a nutritional requirement for essential metals for normal growth and health. But in high doses many of these metals become toxic. Understandably, there is also a need to keep these metals in balance. We are studying the genes that help maintain this equilibrium. The long term goal is to understand how deregulation of these same genes contributes to metal induced disease
Keywords: Address; Affect; Autoregulation; Binding; Binding (Molecular Function); Binding Proteins; Binding Sites; Biological; Biological Models; Brain; Cadmium; Cd element; Cd thionein; Cell Communication and Signaling; Cell Function; Cell Process; Cell Signaling; Cell physiology; Cells; Cellular Function; Cellular Physiology; Cellular Process; Cerebral Pseudosclerosis; Cognitive Discrimination; Combining Site; Complex; Connective Tissue; Copper; Cu element; Discrimination; Discrimination (Psychology); Disease; Disorder; Dose; Drosophila; Drosophila genome; Drosophila genus; Elongation Factor; Encephalon; Encephalons; Equilibrium; Fe element; Free Radicals; Fruit Fly, Drosophila; Gene Action Regulation; Gene Expression Profile; Gene Expression Regulation; Gene Regulation; Gene Regulation Process; Gene Targeting; Gene Transcription; Generalized Growth; Generations; Genes; Genetic Transcription; Genomics; Goals; Gowers` chorea; Growth; Health; Hepato-Neurologic Wilson Disease; Hepatocerebral Degeneration; Hepatolenticular Degeneration; Hg element; Homeostasis; Human; Human, General; Hypocupremia, Congenital; Hypocupremias, Congenital; Immune Precipitation; Immunoprecipitation; Individual; Intracellular Communication and Signaling; Iron; Kidney; Kinky Hair Disease; Kinky Hair Syndrome; Kinnier-Wilson Disease; Knowledge; Lead; Left; Libraries; Ligand Binding Protein; Liver; MRE-binding transcription factor; MTF-1 protein; Mammalian Cell; Man (Taxonomy); Man, Modern; Mediator; Mediator of Activation; Mediator of activation protein; Menkes Kinky Hair Syndrome; Menkes Syndrome; Menkes disease; Menkes kinky hair syndrome (MKHS); Menkes` Disease; Mercury; Metabolic; Metallothionein; Metals; Microarray Analysis; Microarray-Based Analysis; Model System; Modeling; Models, Biologic; Molecular Interaction; NRVS-SYS; Nature; Nervous System; Nervous System, Brain; Nervous system structure; Neurohepatic Degeneration; Neurologic Body System; Neurologic Organ System; Normal Cell; Nutrient; Nutritional Requirements; Organism; Pathology; Pb element; Physiologic; Physiological; Physiological Homeostasis; Play; Position; Positioning Attribute; Post-Transcriptional Gene Silencing; Post-Transcriptional Gene Silencings; Posttranscriptional Gene Silencing; Posttranscriptional Gene Silencings; Programs (PT); Programs [Publication Type]; Progressive Lenticular Degeneration; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Proteins; Pseudosclerosis; Quelling; RNA Expression; RNA Interference; RNA Silencing; RNA Silencings; RNAi; Reactive Site; Regulation; Regulon; Response Elements; Role; Sequence-Specific DNA Binding Protein; Sequence-Specific Posttranscriptional Gene Silencing; Signal Transduction; Signal Transduction Systems; Signaling; Staging; Steely Hair Disease; Steely Hair Syndrome; Stimulus; Stress; Subcellular Process; System; System, LOINC Axis 4; Targetings, Gene; Tissue Growth; Toxic effect; Toxicities; Transcript; Transcription; Transcription Activation; Transcription Elongation; Transcription Initiation; Transcription Regulation; Transcription, Genetic; Transcriptional Activation; Transcriptional Control; Transcriptional Regulation; Transition Elements; Up-Regulation; Urinary System, Kidney; Westphal Strumpell disease; Westphal pseudosclerosis; Westphal-Strumpell Syndrome; Wilson Disease; Work; X-linked copper malabsorption; Zinc; Zn element; balance; balance function; base; biological signal transduction; body system, hepatic; cadmium metallothionein; cadmium-thionein; cohort; congenital hypocupraemia syndrome; copper transport disease; disease/disorder; familial hepatitis; fruit fly; gene expression signature; gene product; genome, Drosophila; genome, fruit fly; heavy metal Pb; heavy metal lead; in vitro Assay; interest; kinky hair syndrome (KHS); living system; metal-responsive transcription factor-1; microarray technology; nutrient requirement; ontogeny; organ system, hepatic; prevent; preventing; programs; public health relevance; renal; response; sensor; sex-linked neurodegenerative disease with monilethrix; social role; toxic metal; transcription factor; transcription factor MTF-1; transcriptome; transition metal; trichopoliodystrophy
Relevance: Humans have a nutritional requirement for essential metals for normal growth and health. But in high doses many of these metals become toxic. Understandably, there is also a need to keep these metals in balance. We are studying the genes that help maintain this equilibrium. The long term goal is to understand how deregulation of these same genes contributes to metal induced disease
Project start date: 2010-09-01
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: PA-07-070
3R01GM085250-02S1 (2010): $35000
1R01GM085250-01A2 (2009): $296596