Activation of eNOS in endothelial cells exposed to ionizing radiation involves components of the DNA damage response pathway

Masaki Nagane; Hironobu Yasui; Yuri Sakai; Tohru Yamamori; Koichi Niwa; Yuichi Hattori; Takashi Kondo; Osamu Inanami

doi:10.1016/j.bbrc.2014.12.002

Activation of eNOS in endothelial cells exposed to ionizing radiation involves components of the DNA damage response pathway

Biochem Biophys Res Commun. 2015 Jan 2;456(1):541-6. doi: 10.1016/j.bbrc.2014.12.002. Epub 2014 Dec 8.

Authors

Masaki Nagane¹, Hironobu Yasui¹, Yuri Sakai¹, Tohru Yamamori¹, Koichi Niwa², Yuichi Hattori³, Takashi Kondo⁴, Osamu Inanami⁵

Affiliations

¹ Laboratory of Radiation Biology, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
² Laboratory of Biochemistry, Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, Abashiri 099-2493, Japan.
³ Department of Molecular and Medical Pharmacology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama 930-0194, Japan.
⁴ Department of Radiological Sciences, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama 930-0194, Japan.
⁵ Laboratory of Radiation Biology, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan. Electronic address: inanami@vetmed.hokudai.ac.jp.

PMID: 25498542
DOI: 10.1016/j.bbrc.2014.12.002

Abstract

In this study, the involvement of ataxia telangiectasia mutated (ATM) kinase and heat shock protein 90 (HSP90) in endothelial nitric oxide synthase (eNOS) activation was investigated in X-irradiated bovine aortic endothelial cells. The activity of nitric oxide synthase (NOS) and the phosphorylation of serine 1179 of eNOS (eNOS-Ser1179) were significantly increased in irradiated cells. The radiation-induced increases in NOS activity and eNOS-Ser1179 phosphorylation levels were significantly reduced by treatment with either an ATM inhibitor (Ku-60019) or an HSP90 inhibitor (geldanamycin). Geldanamycin was furthermore found to suppress the radiation-induced phosphorylation of ATM-Ser1181. Our results indicate that the radiation-induced eNOS activation in bovine aortic endothelial cells is regulated by ATM and HSP90.

Keywords: ATM; DNA damage response; Endothelial nitric oxide synthase; HSP90; Ionizing radiation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Aorta / cytology*
Aorta / radiation effects
Ataxia Telangiectasia Mutated Proteins / metabolism
Benzoquinones / chemistry
Cattle
Cytoplasm / metabolism
DNA Damage*
DNA Repair*
Endothelial Cells / cytology
Endothelial Cells / radiation effects*
Gene Expression Regulation, Enzymologic*
HSP90 Heat-Shock Proteins / metabolism
Immunohistochemistry
Lactams, Macrocyclic / chemistry
Morpholines / chemistry
Nitric Oxide Synthase / metabolism
Nitric Oxide Synthase Type III / metabolism*
Phosphorylation
Radiation, Ionizing
Thioxanthenes / chemistry
Time Factors
X-Rays

Substances

2-(2,6-dimethylmorpholin-4-yl)-N-(5-(6-morpholin-4-yl-4-oxo-4H-pyran-2-yl)-9H-thioxanthen-2-yl)acetamide
Benzoquinones
HSP90 Heat-Shock Proteins
Lactams, Macrocyclic
Morpholines
Thioxanthenes
Nitric Oxide Synthase
Nitric Oxide Synthase Type III
Ataxia Telangiectasia Mutated Proteins
geldanamycin