Structure and regulation of the human muscle-specific enolase gene
Structure and regulation of the human muscle-specific enolase gene
The enzyme enolase (E.C.4.2.1.11), catalyses the interconversion of 2-phosphoglycerate to phosphoenolpyruvate and occurs midway through the glycolytic pathway. Mammals and birds exhibit three specific isoenzymes; muscle-specific enolase [MSE] expressed in adult skeletal muscle, neuron-specific enolase (NSE) expressed primarily in neurons and non-neuronal enolase (NNE) expressed in foetal and other cell types. This thesis describes the characterisation of the human MSE gene and the studies of the transcriptional regulation of this gene. Chapter 1 reviews the current literature on MSE with respect to the other two isoenzymes, NSE and NNE and includes a brief summary of the aims of the project. Chapter 2 outlines the general methodology used throughout the project. Chapter 3 describes the characterisation of the human MSE gene. Its exon-intron structure is similar to that of the other isoforms. The 5' end displays features characteristic of a CpG-rich island. In keeping with its role as a muscle-specific gene, three muscle-specific regulatory elements, M-CAT, MEF-1 and CArG were identified. This study also demonstrates that the MSE transcript is expressed specifically in adult skeletal muscle. Chapter 4 describes methylation studies carried out to determine the role of DNA methylation in the regulation of MSE gene expression. The studies show that the 4kb CpG-rich island is methylation-free. One site however, in intron 6 is partially demethylated in skeletal muscle but fully methylated in sperm and brain. The implications of the methylation-free island and the site-specific demethylation are discussed. Chapter 5 describes the characterisation of the MSE promoter, by transient transfection studies and CAT assays in a mouse muscle cell line. Although the promoter region per se is not able to drive CAT gene expression, proximal and distal domains of the promoter region display 20-30 fold higher CAT activity equally, at both the myoblast and myotube stage. The role of the MSE gene as an early marker of gene expression, and the potential function of the muscle-specific regulatory elements are discussed. Chapter 6 presents a general discussion of the work carried out for this project and offers possible future directions for further investigations.
University of Southampton
1991
Peshavaria, Mina
(1991)
Structure and regulation of the human muscle-specific enolase gene.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
The enzyme enolase (E.C.4.2.1.11), catalyses the interconversion of 2-phosphoglycerate to phosphoenolpyruvate and occurs midway through the glycolytic pathway. Mammals and birds exhibit three specific isoenzymes; muscle-specific enolase [MSE] expressed in adult skeletal muscle, neuron-specific enolase (NSE) expressed primarily in neurons and non-neuronal enolase (NNE) expressed in foetal and other cell types. This thesis describes the characterisation of the human MSE gene and the studies of the transcriptional regulation of this gene. Chapter 1 reviews the current literature on MSE with respect to the other two isoenzymes, NSE and NNE and includes a brief summary of the aims of the project. Chapter 2 outlines the general methodology used throughout the project. Chapter 3 describes the characterisation of the human MSE gene. Its exon-intron structure is similar to that of the other isoforms. The 5' end displays features characteristic of a CpG-rich island. In keeping with its role as a muscle-specific gene, three muscle-specific regulatory elements, M-CAT, MEF-1 and CArG were identified. This study also demonstrates that the MSE transcript is expressed specifically in adult skeletal muscle. Chapter 4 describes methylation studies carried out to determine the role of DNA methylation in the regulation of MSE gene expression. The studies show that the 4kb CpG-rich island is methylation-free. One site however, in intron 6 is partially demethylated in skeletal muscle but fully methylated in sperm and brain. The implications of the methylation-free island and the site-specific demethylation are discussed. Chapter 5 describes the characterisation of the MSE promoter, by transient transfection studies and CAT assays in a mouse muscle cell line. Although the promoter region per se is not able to drive CAT gene expression, proximal and distal domains of the promoter region display 20-30 fold higher CAT activity equally, at both the myoblast and myotube stage. The role of the MSE gene as an early marker of gene expression, and the potential function of the muscle-specific regulatory elements are discussed. Chapter 6 presents a general discussion of the work carried out for this project and offers possible future directions for further investigations.
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Published date: 1991
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Local EPrints ID: 460645
URI: http://eprints.soton.ac.uk/id/eprint/460645
PURE UUID: e01d1a86-6c12-4d3d-a11d-e1ecb5101255
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Date deposited: 04 Jul 2022 18:26
Last modified: 04 Jul 2022 18:26
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Author:
Mina Peshavaria
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