Molecular Characterization of the Proximal Human Endothelial Nitric Oxide Synthase (eNOS) Promoter PDF Download

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Languages : en
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Author: Fotula Karantzoulis-Fegaras
Publisher: National Library of Canada = Bibliothèque nationale du Canada
ISBN: 9780612457331
Category :
Languages : en
Pages : 548

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Author: Fotula Karantzoulis-Fegaras
Publisher:
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Category :
Languages : en
Pages : 0

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Understanding transcription initiation of the endothelial nitric oxide synthase (eNOS) gene appears pivotal to gaining a comprehensive view of NO biology in the blood vessel wall. The present thesis focused upon a detailed dissection of the functionally important cis-DNA elements and the multiprotein complexes implicated in the cooperative control of the constitutive expression of the human eNOS gene in vascular endothelium. Three tightly clustered cis-regulatory regions were identified in the proximal enhancer of the TATA-less eNOS promoter using deletion analysis and linker-scanning mutagenesis: positive regulatory domain I (PRD I) -104/-95 relative to transcription initiation), PRD II (-144/-115) and PRD III (-44/-25). The nucleoprotein complexes that form upon these regions in endothelial cells contained Ets family members, Sp1 and variants of Sp3, MAZ and YY1. Functional domain studies in 'Drosophila' Schneider cells and endothelial cells revealed examples of positive and negative protein-protein cooperativity involving Sp1, variants of Sp3, Ets-1, Elf-1 and MAZ. The multiprotein complexes formed on the activator recognition sites within this region of the human eNOS promoter identify the existence of a constitutively active enhanceosome in vascular endothelium. Moreover, 'in vitro' DNase I footprinting analyses and nuclear run-off studies also indicate their important contributions to preinitiation complex formation. Surprisingly, episomal expression studies performed in endothelial and non-endothelial cell types revealed that eNOS promoter/reporter constructs lacked cell-specificity, suggesting that there exist cell-specific mechanisms that repress native eNOS gene expression in non-expressing cell types. We posited that DNA methylation played a role in this cell-specific expression. Sodium bisulfite genomic sequencing revealed a different methylation pattern of the eNOS promoter in endothelial versus non-endothelial cells. Transient transfection of 'in vitro' methylated promoter/reporter constructs revealed a possible role for methylation in transcriptional repression. DNA hypomethylation studies performed with 5-azacytidine, resulted in increased expression of eNOS mRNA in cell types that do not normally express the eNOS mRNA transcript. The role of epigenetic regulation in cell-specific eNOS gene expression is a novel finding in the vascular endothelium. We have identified novel concepts with respect to protein-DNA interactions, pre-initiation complex formation and chromatin-based epigenetic mechanisms in vascular endothelium.

Author: Terrie K. Rife
Publisher:
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Category :
Languages : en
Pages : 324

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Author: Michel Félétou
Publisher: Morgan & Claypool Publishers
ISBN: 1615041230
Category : Science
Languages : en
Pages : 309

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The endothelium, a monolayer of endothelial cells, constitutes the inner cellular lining of the blood vessels (arteries, veins and capillaries) and the lymphatic system, and therefore is in direct contact with the blood/lymph and the circulating cells. The endothelium is a major player in the control of blood fluidity, platelet aggregation and vascular tone, a major actor in the regulation of immunology, inflammation and angiogenesis, and an important metabolizing and an endocrine organ. Endothelial cells controls vascular tone, and thereby blood flow, by synthesizing and releasing relaxing and contracting factors such as nitric oxide, metabolites of arachidonic acid via the cyclooxygenases, lipoxygenases and cytochrome P450 pathways, various peptides (endothelin, urotensin, CNP, adrenomedullin, etc.), adenosine, purines, reactive oxygen species and so on. Additionally, endothelial ectoenzymes are required steps in the generation of vasoactive hormones such as angiotensin II. An endothelial dysfunction linked to an imbalance in the synthesis and/or the release of these various endothelial factors may explain the initiation of cardiovascular pathologies (from hypertension to atherosclerosis) or their development and perpetuation. Table of Contents: Introduction / Multiple Functions of the Endothelial Cells / Calcium Signaling in Vascular Cells and Cell-to-Cell Communications / Endothelium-Dependent Regulation of Vascular Tone / Conclusion / References

Author: Ismail Laher
Publisher: Springer
ISBN: 9783642300172
Category : Medical
Languages : en
Pages : 0

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The focus of this collection of illustrated reviews is to discuss the systems biology of free radicals and anti-oxidants. Free radical induced cellular damage in a variety of tissues and organs is reviewed, with detailed discussion of molecular and cellular mechanisms. The collection is aimed at those new to the field, as well as clinicians and scientists with long standing interests in free radical biology. A feature of this collection is that the material also brings insights into various diseases where free radicals are thought to play a role. There is extensive discussion of the success and limitations of the use of antioxidants in several clinical settings.

Author: Ernst van Faassen
Publisher: Elsevier
ISBN: 0080489591
Category : Science
Languages : en
Pages : 444

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Radicals for Life: the Various Forms of Nitric Oxide provides an up-to-date overview of the role of nitrosocompounds and nitrosyl-iron complexes in physiology. Nitrosocompounds can be considered as stabilised forms of nitric oxide, one of the most important regulatory molecules in physiology today. Many nitrosocompounds share some of the physiological functions of nitric oxide, and may be formed inside living organisms. This is the first book to be published that is dedicated to the role of such nitrosocompounds in physiology, with particular emphasis on the nitrosocompounds that are endogenously formed in higher organisms and humans. Points of discussion include: physical and chemical properties of the compounds, the main chemical pathways in vivo, as well as the physiological effects that have been recognised to date. Each of the nineteen chapters is written by distinguished specialists in the field, well known for their original and important contributions to the subject. Also included are results from a wide range of studies in vitro, in cell cultures, animal models and human volunteers. Examples of alternative forms of nitric oxide, with special emphasis on their protective role against widespread human diseases like atherosclerosis, Alzheimer's disease, diabetes, sexual dysfunction, and renal insufficiency to stroke and ischemia are also included. First monograph to consider and provide an overview of endogenous nitrosocompounds and nitrosyl-iron complexes Extensive bibliographic references, written by specialists of human physiology Providing high scientific quality with a focus on implications for human diseases

Author: Helena Lenasi
Publisher: BoD – Books on Demand
ISBN: 1789842530
Category : Medical
Languages : en
Pages : 432

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The endothelium enables communication between blood and tissues and is actively involved in cardiovascular homeostasis. Endothelial dysfunction has been recognized as an early step in the development of cardiovascular diseases: respectively, endothelium represents a potential therapeutic niche with multiple targets. The purpose of the book is to point out some recent findings of endothelial physiology and pathophysiology emphasizing various aspects of endothelial dysfunction connected to the body's internal and external environment. While basic features of the endothelium are presented in an introductory chapter, the authors of the following 17 chapters have provided extensive insight into some selected topics of endothelial (dys)function. The book would hopefully be useful for anyone interested in recapitulating endothelial (patho)physiology and expanding knowledge of molecular mechanisms involved in endothelial dysfunction, relevant also for further clinical investigations.

Author:
Publisher: Academic Press
ISBN: 0080536379
Category : Science
Languages : en
Pages : 375

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Although the function of nitric oxide in a regulatory capacity in the central and peripheral nervous system is widely recognized, the full scope of its actions and its interrelationships with other classes of regulatory molecules is just beginning to be comprehended. This volume contains a number of sophisticated and advanced methods essential for exploring the activity of nitric oxide in the brain. It will be a valuable tool for the established investigator and for those just entering the field. Comprehensive protocols included for detection of NO and related compounds by chemical, immunohistochemical, and in situ hybridization techniques Newly developed methods for the purification of neuronal and endothelial NO synthase, production of monoclonal antibodies to NO synthase, molecular cloning and expression of NO synthesis, and control of NO synthase gene expression Assessment of NO-mediated functions in neurons, central nervous system, cerebral circulation, synaptic transmission, and vascular tone Calcium imaging by confocal microscopy, evaluation of the effect of NO on iron metabolism, and detection of heme oxygenase-1 and -2 message level and distribution

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ISBN:
Category : Dissertation abstracts
Languages : en
Pages : 848

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