GTP cyclohydrolase I expression is regulated by nitric oxide: Role of cyclic AMP

Sanjiv Kumar, Xutong Sun, Shruti Sharma, Saurabh Aggarwal, Kandasamy Ravi, Jeffery R. Fineman, Stephen M. Black

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Our previous studies have demonstrated that nitric oxide (NO) leads to nitric oxide synthase (NOS) uncoupling and an increase in NOSderived superoxide. However, the cause of this uncoupling has not been adequately resolved. The pteridine cofactor tetrahydrobiopterin (BH4) is a critical determinant of endothelial NOS (eNOS) activity and coupling, and GTP cyclohydrolase I (GCH1) is the rate-limiting enzyme in its generation. Thus the initial purpose of this study was to determine whether decreases in BH 4 could underlie, at least in part, the NO-mediated uncoupling of eNOS we have observed both in vitro and in vivo. Initially we evaluated the effect of inhaled NO levels on GCH1 expression and BH4 levels in the intact lamb. Contrary to our hypothesis, we found that there was a significant increase in both plasma BH4 levels and peripheral lung GCH1 protein levels. Furthermore, in vitro, we found that exposure to the NO donor spermine NONOate (SPNONO) led to an increase in GCH1 protein and BH4 levels in both COS-7 and pulmonary arterial endothelial cells. However, SPNONO treatment also caused a significant increase in phospho-cAMP response element binding protein (CREB) levels, as detected by Western blot analysis, and significantly increased cAMP levels, as detected by enzyme immunoassay. Furthermore, utilizing GCH1 promoter fragments fused to a luciferase reporter gene, we found that GCH1 promoter activity was enhanced by SPNONO in a CREB-dependent manner, and electromobility shift assays revealed an NO-dependent increase in the nuclear binding of CREB. These data suggest that NO increases BH4 levels through a cAMP/CREBmediated increase in GCH1 transcription and that the eNOS uncoupling associated with exogenous NO does not involved reduced BH4 levels.

Original languageEnglish (US)
Pages (from-to)L309-L317
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume297
Issue number2
DOIs
StatePublished - Aug 1 2009

Keywords

  • Adenosine 3′,5′-cyclic monophosphate response element binding protein
  • Cell signaling
  • Rebound pulmonary hypertension
  • Tetrahydrobiopterin

ASJC Scopus subject areas

  • Physiology
  • Pulmonary and Respiratory Medicine
  • Physiology (medical)
  • Cell Biology

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