Evidence That E-Box Promoter Elements and MyoD Transcription Factors Play a Role in the Induction of Cathepsin B Gene Expression during Human Myoblast Differentiation
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D.T. Jane
Abstract
HB13 human myoblasts express physiological and biochemical markers associated with myoblast differentiation in nonhuman cell culture model systems. During differentiation, HB13 myoblasts also demonstrate fusionrelated increases in cathepsin B activity and protein levels. These increases are associated with an increase in levels of cathepsin B mRNA suggesting the involvement of transcriptional regulatory mechanisms. To examine these mechanisms human myoblasts were transfected with cathepsin B nested deletion promoter constructs within the 1.8 kb 5 promoter 1 region of the human catB gene. Transfected myoblasts that were maintained under differentiating conditions demonstrated higher promoter activity than those maintained in proliferating conditions. The highest activity was obtained with pSCB2-3 ( 1279/ +56 bp), a construct containing two putative upstream Ebox elements. Cotransfection experiments demonstrated that MyoD and myogenin transactivate cathepsin B promoter activity. Electrophoretic mobility shift assays of nuclear extracts incubated with an oligonucleotide containing two upstream Ebox elements found within the cathepsin B promoter demonstrated two band shifts. The band shifts were abolished using an oligonucleotide with mutations in both Ebox elements. Moreover, the shifted bands were supershifted and abolished when incubated with antimyogenin and antiMyoD, respectively. Collectively, these data support myogenic transcription factor mediated activation of cathepsin B expression during myogenesis.
Copyright © 2002 by Walter de Gruyter GmbH & Co. KG
Abstract
HB13 human myoblasts express physiological and biochemical markers associated with myoblast differentiation in nonhuman cell culture model systems. During differentiation, HB13 myoblasts also demonstrate fusionrelated increases in cathepsin B activity and protein levels. These increases are associated with an increase in levels of cathepsin B mRNA suggesting the involvement of transcriptional regulatory mechanisms. To examine these mechanisms human myoblasts were transfected with cathepsin B nested deletion promoter constructs within the 1.8 kb 5 promoter 1 region of the human catB gene. Transfected myoblasts that were maintained under differentiating conditions demonstrated higher promoter activity than those maintained in proliferating conditions. The highest activity was obtained with pSCB2-3 ( 1279/ +56 bp), a construct containing two putative upstream Ebox elements. Cotransfection experiments demonstrated that MyoD and myogenin transactivate cathepsin B promoter activity. Electrophoretic mobility shift assays of nuclear extracts incubated with an oligonucleotide containing two upstream Ebox elements found within the cathepsin B promoter demonstrated two band shifts. The band shifts were abolished using an oligonucleotide with mutations in both Ebox elements. Moreover, the shifted bands were supershifted and abolished when incubated with antimyogenin and antiMyoD, respectively. Collectively, these data support myogenic transcription factor mediated activation of cathepsin B expression during myogenesis.
Copyright © 2002 by Walter de Gruyter GmbH & Co. KG
Articles in the same Issue
- Processing and Activation of Lysosomal Proteinases
- Evidence That E-Box Promoter Elements and MyoD Transcription Factors Play a Role in the Induction of Cathepsin B Gene Expression during Human Myoblast Differentiation
- Transcription Factor Egr-1 Activates Collagen Expression in Immortalized Fibroblasts or Fibrosarcoma Cells
- The Receptor-Bound N-Terminal Ectodomain of the Amyloid Precursor Protein Is Associated with Membrane Rafts
- Evaluation of Similarities in the cis/trans Isomerase Function of Trigger Factor and DnaK
- The Chloroplast Import Receptor Toc34 Functions as Preprotein-Regulated GTPase
- cis-4-Methylsphingosine Phosphate Induces Apoptosis in Neuroblastoma Cells by Opposite Effects on p38 and ERK Mitogen-Activated Protein Kinases
- Evidence for the Involvement of the Unique C-Tail of S100A9 in the Binding of Arachidonic Acid to the Heterocomplex S100A8/A9
- The Primary Structure of Three Hemoglobin Chains from the Indigo Snake (Drymarchon corais erebennus, Serpentes): First Evidence for αD Chains and Two β Chain Types in Snakes
- Expression of the Tissue Kallikrein-Kinin Cascade in Granulosa Cells of the Ovary
- Chondroitin Sulfate Proteoglycan Is a Potent Enhancer in the Processing of Procathepsin L
- Tyrosinase-Mediated Oxidation of Acetaminophen to 4-Acetamido-o- Benzoquinone
- Properties of Baculovirus Particles Displaying GFP Analyzed by Fluorescence Correlation Spectroscopy
- The Promoter Context Determines Mutual Repression or Synergism between NF-κB and the Glucocorticoid Receptor
- Expression of Hepatocyte Growth Factor Activator Inhibitor Type 2 (HAI-2) in Human Testis: Identification of a Distinct Transcription Start Site for the HAI-2 Gene in Testis
- Identification and Characterization of an IgG Binding Protein in the Secretion of the Rat Coagulating Gland
- Characterization of Different Reactive Lysines in Bovine Heart Mitochondrial Porin
- Determination of the mRNA Sequence of Cathepsin Y, a Cysteine Endopeptidase from Rat Spleen, and Confirmation of Its Ubiquitous
- Acknowledgement
- Content Index
- Author Index
- Subject Index
Articles in the same Issue
- Processing and Activation of Lysosomal Proteinases
- Evidence That E-Box Promoter Elements and MyoD Transcription Factors Play a Role in the Induction of Cathepsin B Gene Expression during Human Myoblast Differentiation
- Transcription Factor Egr-1 Activates Collagen Expression in Immortalized Fibroblasts or Fibrosarcoma Cells
- The Receptor-Bound N-Terminal Ectodomain of the Amyloid Precursor Protein Is Associated with Membrane Rafts
- Evaluation of Similarities in the cis/trans Isomerase Function of Trigger Factor and DnaK
- The Chloroplast Import Receptor Toc34 Functions as Preprotein-Regulated GTPase
- cis-4-Methylsphingosine Phosphate Induces Apoptosis in Neuroblastoma Cells by Opposite Effects on p38 and ERK Mitogen-Activated Protein Kinases
- Evidence for the Involvement of the Unique C-Tail of S100A9 in the Binding of Arachidonic Acid to the Heterocomplex S100A8/A9
- The Primary Structure of Three Hemoglobin Chains from the Indigo Snake (Drymarchon corais erebennus, Serpentes): First Evidence for αD Chains and Two β Chain Types in Snakes
- Expression of the Tissue Kallikrein-Kinin Cascade in Granulosa Cells of the Ovary
- Chondroitin Sulfate Proteoglycan Is a Potent Enhancer in the Processing of Procathepsin L
- Tyrosinase-Mediated Oxidation of Acetaminophen to 4-Acetamido-o- Benzoquinone
- Properties of Baculovirus Particles Displaying GFP Analyzed by Fluorescence Correlation Spectroscopy
- The Promoter Context Determines Mutual Repression or Synergism between NF-κB and the Glucocorticoid Receptor
- Expression of Hepatocyte Growth Factor Activator Inhibitor Type 2 (HAI-2) in Human Testis: Identification of a Distinct Transcription Start Site for the HAI-2 Gene in Testis
- Identification and Characterization of an IgG Binding Protein in the Secretion of the Rat Coagulating Gland
- Characterization of Different Reactive Lysines in Bovine Heart Mitochondrial Porin
- Determination of the mRNA Sequence of Cathepsin Y, a Cysteine Endopeptidase from Rat Spleen, and Confirmation of Its Ubiquitous
- Acknowledgement
- Content Index
- Author Index
- Subject Index
