Graduation Year
2005
Document Type
Dissertation
Degree
Ph.D.
Degree Granting Department
Biochemistry and Molecular Biology
Major Professor
Duane C. Eichler, Ph.D.
Committee Member
Larry P. Solomonson, Ph.D.
Committee Member
Denise Cooper, Ph.D.
Committee Member
Huntington Potter, Ph.D.
Committee Member
Ken Wright, Ph.D.
Keywords
Nitric oxide, Enos, Endothelial, Apoptosis, tnf-alpha, ppar-gamma
Abstract
While cellular levels of arginine greatly exceed the apparent Km for endothelial nitric oxide synthase (eNOS), nitric oxide (NO) production is limited by availability of arginine. Results from this work have provided a unique understanding of endothelial NO production, showing that arginine regeneration, that is the recycling of citrulline back to arginine by argininosuccinate synthase (AS) and argininosuccinate lyase (AL), defines the essential source of arginine for NO production. Using RNA interference analysis, selective reduction of AS expression was shown to directly correspond with a diminished capacity of endothelial cells to produce NO, despite saturating levels of arginine in the medium. In addition, the viability of AS siRNA-treated endothelial cells was compromised due to apoptotic cell death.
AS expression was also investigated in response to two major vascular effectors. Tumor necrosis factor (TNF)-α, which is known to impair endothelial NO production, was shown to provoke a dose-dependent reduction of AS expression that corresponded to a decrease in NO production. Furthermore, TNF-α was shown to suppress AS expression through a NFκB mediated pathway, which involves three essential Sp1 elements in the proximal AS gene promoter. On the other hand, peroxisome proliferator-activated receptor gamma (PPARγ) agonists, troglitazone and ciglitazone, which are known to elicit a vascular protective response against TNF-α effects, were shown to coordinately induce NO production and AS expression via a PPARγ response element in the distal AS gene promoter. Importantly, these PPARγ agonists were shown to restore AS expression and NO production following down-regulation by TNF-α, consistent with their vascular protective properties.
Scholar Commons Citation
Goodwin, Bonnie L., "The Role and Regulation of Argininosuccinate Synthase in Endothelial Function" (2005). USF Tampa Graduate Theses and Dissertations.
https://digitalcommons.usf.edu/etd/2902
