Evidence That an Endothelial Cytosolic Protein Binds to the 3′-Untranslated Region of Endothelial Nitric Oxide Synthase mRNA

Abstract

Changes in the endothelial nitric oxide synthase (eNOS) expression could be involved in the endothelium-dependent vasorelaxing dysfunction associated with cardiovascular diseases. We have recently demonstrated the existence of endothelial cytosolic proteins that bind to the 3′-untranslated region (3′-UTR) of eNOS mRNA and could be involved in eNOS mRNA stabilization. In the present work, we have characterized the cytosolic proteins that bind to 3′-UTR eNOS mRNA. An endothelial cytosolic protein (MW 60-kD) specifically bound to 3′-UTR eNOS mRNA as determined by a cross-linking assay followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The endothelial cytosolic protein recognized a cytidine (C)-rich region within 3′-UTR eNOS mRNA. Furthermore, tumor necrosis factor-α (TNF-α) increased the level of the 60-kD endothelial cytosolic protein. In addition, TNF-α reduced eNOS mRNA levels and this was prevented by coincubation with cycloheximide. Cycloheximide also prevented the binding activity of the endothelial cytosolic protein to 3′-UTR eNOS mRNA. In summary, these data suggest that a 60-kD endothelial cytosolic protein binds to 3′-UTR eNOS mRNA. TNF-α increased the 60-kD protein levels. Cycloheximide prevented the binding activity of the cytosolic protein to 3′-UTR eNOS mRNA related to TNF-α; this effect was associated with greater eNOS mRNA levels. Further specific studies are needed to determine the involvement of this 60-kD endothelial cytosolic protein in the regulation of eNOS mRNA stabilization and in the endothelial dysfunction associated with cardiovascular diseases.

Journal

Journal ID (publisher-id): JVR

Journal ID (nlm-ta): J Vasc Res

Journal ID (doi): 10.1159/issn.1018-1172

Title: Journal of Vascular Research

Publisher: S. Karger AG

ISSN (Print): 1018-1172

ISSN (Electronic): 1423-0135

Publication date Subscription-year: 1999

Publication date (Print): June 1999

Publication date (Electronic): 18 June 1999

Volume: 36

Issue: 3

Pages: 201-208

Affiliations

^aNephrology, Hypertension and Cardiovascular Research Laboratory, Fundación Jiménez Díaz, Madrid and ^bDepartment of Biochemistry and Cellular and Molecular Biology, Faculty of Veterinary Medicine, University of Zaragoza, Zaragoza, Spain

Article

Publisher ID: 25643 Other ID: J Vasc Res 1999;36:201–208

DOI: 10.1159/000025643

PubMed ID: 10393506

Copyright statement: © 1999 S. Karger AG, Basel

License:

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Page count

Figures: 7, References: 25, Pages: 8

Product

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Categories

Subject: Research Paper