TY - JOUR
T1 - Nitric oxide regulates pulmonary vascular smooth muscle cell expression of the inducible cAMP early repressor gene
AU - Steinbicker, Andrea U.
AU - Liu, Heling
AU - Jiramongkolchai, Kim
AU - Malhotra, Rajeev
AU - Choe, Elizabeth Y.
AU - Busch, Cornelius J.
AU - Graveline, Amanda R.
AU - Kao, Sonya M.
AU - Nagasaka, Yasuko
AU - Ichinose, Fumito
AU - Buys, Emmanuel S.
AU - Brouckaert, Peter
AU - Zapol, Warren M.
AU - Bloch, Kenneth D.
N1 - Funding Information:
KDB receives research Grant support via a sponsored research agreement with INOTherapeutics and the Massachusetts General Hospital. WMZ receives royalties on patents licensed by Massachusetts General Hospital to Linde Corp (Munich, Germany) and Ikaria (Clinton, New Jersey) on inhaled NO.
Funding Information:
This work was supported by the German DFG grant SW 119-3/1 (AUS), a Scientist Development Grant 9930085N from the American Heart Association (HL), a Research Training Fellowship for Medical Students from the Howard Hughes Medical Institute (KJ), NHLBI T32HL007208 (RM), a Scientist Development Grant 10SDG2610313 from the American Heart Association (ESB), grants from the FWO-Vlaanderen and the UGent-GOA programs (PB), as well as grants from the Department of Anesthesia at MGH (WMZ) and grants from the NHLBI, HL-57172 and R01 HL074352 (KDB).
PY - 2011/10/30
Y1 - 2011/10/30
N2 - Nitric oxide (NO) regulates vascular smooth muscle cell (VSMC) structure and function, in part by activating soluble guanylate cyclase (sGC) to synthesize cGMP. The objective of this study was to further characterize the signaling mechanisms by which NO regulates VSMC gene expression using transcription profiling. DNA microarrays were hybridized with RNA extracted from rat pulmonary artery smooth muscle cells (RPaSMC) exposed to the NO donor compound, S-nitroso-glutathione (GSNO). Many of the genes, whose expression was induced by GSNO, contain a cAMP-response element (CRE), of which one encoded the inducible cAMP early repressor (ICER). sGC and cAMP-dependent protein kinase, but not cGMP-dependent protein kinase, were required for NO-mediated phosphorylation of CRE-binding protein (CREB) and induction of ICER gene expression. Expression of a dominant-negative CREB in RPaSMC prevented the NO-mediated induction of CRE-dependent gene transcription and ICER gene expression. Pre-treatment of RPaSMC with the intracellular calcium (Ca 2+) chelator, BAPTA-AM, blocked the induction of ICER gene expression by GSNO. The store-operated Ca 2+ channel inhibitors, 2-ABP, and SKF-96365, reduced the GSNO-mediated increase in ICER mRNA levels, while 2-ABP did not inhibit GSNO-induced CREB phosphorylation. Our results suggest that induction of ICER gene expression by NO requires both CREB phosphorylation and Ca 2+ signaling. Transcription profiling of RPaSMC exposed to GSNO revealed important roles for sGC, PKA, CREB, and Ca 2+ in the regulation of gene expression by NO. The induction of ICER in GSNO-treated RPaSMC highlights a novel cross-talk mechanism between cGMP and cAMP signaling pathways.
AB - Nitric oxide (NO) regulates vascular smooth muscle cell (VSMC) structure and function, in part by activating soluble guanylate cyclase (sGC) to synthesize cGMP. The objective of this study was to further characterize the signaling mechanisms by which NO regulates VSMC gene expression using transcription profiling. DNA microarrays were hybridized with RNA extracted from rat pulmonary artery smooth muscle cells (RPaSMC) exposed to the NO donor compound, S-nitroso-glutathione (GSNO). Many of the genes, whose expression was induced by GSNO, contain a cAMP-response element (CRE), of which one encoded the inducible cAMP early repressor (ICER). sGC and cAMP-dependent protein kinase, but not cGMP-dependent protein kinase, were required for NO-mediated phosphorylation of CRE-binding protein (CREB) and induction of ICER gene expression. Expression of a dominant-negative CREB in RPaSMC prevented the NO-mediated induction of CRE-dependent gene transcription and ICER gene expression. Pre-treatment of RPaSMC with the intracellular calcium (Ca 2+) chelator, BAPTA-AM, blocked the induction of ICER gene expression by GSNO. The store-operated Ca 2+ channel inhibitors, 2-ABP, and SKF-96365, reduced the GSNO-mediated increase in ICER mRNA levels, while 2-ABP did not inhibit GSNO-induced CREB phosphorylation. Our results suggest that induction of ICER gene expression by NO requires both CREB phosphorylation and Ca 2+ signaling. Transcription profiling of RPaSMC exposed to GSNO revealed important roles for sGC, PKA, CREB, and Ca 2+ in the regulation of gene expression by NO. The induction of ICER in GSNO-treated RPaSMC highlights a novel cross-talk mechanism between cGMP and cAMP signaling pathways.
KW - Cyclic GMP
KW - Nitric oxide
KW - Protein kinase A
KW - Vascular smooth muscle
KW - cAMP-response element
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U2 - 10.1016/j.niox.2011.05.006
DO - 10.1016/j.niox.2011.05.006
M3 - Article
C2 - 21642009
AN - SCOPUS:80053921752
SN - 1089-8603
VL - 25
SP - 294
EP - 302
JO - Nitric Oxide - Biology and Chemistry
JF - Nitric Oxide - Biology and Chemistry
IS - 3
ER -