p90 RSK-1 associates with and inhibits neuronal nitric oxide synthase

Tao Song, Katsuyoshi Sugimoto, Hideshi Ihara, Akihiro Mizutani, Naoya Hatano, Kodai Kume, Toshie Kambe, Fuminori Yamaguchi, Masaaki Tokuda, Yasuo Watanabe

Research output: Contribution to journalArticle

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Abstract

Evidence is presented that RSK1 (ribosomal S6 kinase 1), a downstream target of MAPK (mitogen-activated protein kinase), directly phosphorylates nNOS (neuronal nitric oxide synthase) on Ser847 in response to mitogens. The phosphorylation thus increases greatly following EGF (epidermal growth factor) treatment of rat pituitary tumour GH3 cells and is reduced by exposure to the MEK (MAPK/extracellular-signal-regulated kinase kinase) inhibitor PD98059. Furthermore, it is significantly enhanced by expression of wild-type RSK1 and antagonized by kinase-inactive RSK1 or specific reduction of endogenous RSK1. EGF treatment of HEK-293 (human embryonic kidney) cells, expressing RSK1 and nNOS, led to inhibition of NOS enzyme activity, associated with an increase in phosphorylation of nNOS at Ser847, as is also the case in an in vitro assay. In addition, these phenomena were significantly blocked by treatment with the RSK inhibitor Ro31-8220. Cells expressing mutant nNOS (S847A) proved resistant to phosphorylation and decrease of NOS activity. Within minutes of adding EGF to transfected cells, RSK1 associated with nNOS and subsequently dissociated following more prolonged agonist stimulation. EGF-induced formation of the nNOS-RSK1 complex was significantly decreased by PD98059 treatment. Treatment with EGF further revealed phosphorylation of nNOS on Ser847 in rat hippocampal neurons and cerebellar granule cells. This EGF-induced phosphorylation was partially blocked by PD98059 and Ro31-8220. Together, these data provide substantial evidence that RSK1 associates with and phosphorylates nNOS on Ser847 following mitogen stimulation and suggest a novel role for RSK1 in the regulation of nitric oxide function in brain.

Original languageEnglish
Pages (from-to)391-398
Number of pages8
JournalBiochemical Journal
Volume401
Issue number2
DOIs
Publication statusPublished - Jan 15 2007
Externally publishedYes

Fingerprint

Ribosomal Protein S6 Kinases
Nitric Oxide Synthase Type I
Phosphorylation
Epidermal Growth Factor
Mitogen-Activated Protein Kinases
Mitogens
Rats
Phosphotransferases
Enzyme inhibition
Mitogen-Activated Protein Kinase Kinases
Extracellular Signal-Regulated MAP Kinases
Enzyme activity
Pituitary Neoplasms
Neurons
Tumors
Assays
Brain
Nitric Oxide
Cells
Kidney

Keywords

  • Neuronal nitric oxide synthase
  • Phosphorylation
  • Pituitary tumour GH3 cell
  • Ribosomal S6 kinase

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Song, T., Sugimoto, K., Ihara, H., Mizutani, A., Hatano, N., Kume, K., ... Watanabe, Y. (2007). p90 RSK-1 associates with and inhibits neuronal nitric oxide synthase. Biochemical Journal, 401(2), 391-398. https://doi.org/10.1042/BJ20060580

p90 RSK-1 associates with and inhibits neuronal nitric oxide synthase. / Song, Tao; Sugimoto, Katsuyoshi; Ihara, Hideshi; Mizutani, Akihiro; Hatano, Naoya; Kume, Kodai; Kambe, Toshie; Yamaguchi, Fuminori; Tokuda, Masaaki; Watanabe, Yasuo.

In: Biochemical Journal, Vol. 401, No. 2, 15.01.2007, p. 391-398.

Research output: Contribution to journalArticle

Song, T, Sugimoto, K, Ihara, H, Mizutani, A, Hatano, N, Kume, K, Kambe, T, Yamaguchi, F, Tokuda, M & Watanabe, Y 2007, 'p90 RSK-1 associates with and inhibits neuronal nitric oxide synthase', Biochemical Journal, vol. 401, no. 2, pp. 391-398. https://doi.org/10.1042/BJ20060580
Song, Tao ; Sugimoto, Katsuyoshi ; Ihara, Hideshi ; Mizutani, Akihiro ; Hatano, Naoya ; Kume, Kodai ; Kambe, Toshie ; Yamaguchi, Fuminori ; Tokuda, Masaaki ; Watanabe, Yasuo. / p90 RSK-1 associates with and inhibits neuronal nitric oxide synthase. In: Biochemical Journal. 2007 ; Vol. 401, No. 2. pp. 391-398.
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