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J. Biol. Chem., Vol. 277, Issue 18, 15400-15406, May 3, 2002
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From the Ionizing radiation at clinical dose levels
activates both pro- and anti-proliferative signal transduction
pathways, the balance of which determines cell fate. The initiating and
amplifying mechanisms involved in the activation are poorly understood.
We demonstrate that one mechanism involves stimulation of constitutive
nitric-oxide synthase (NOS) activity. NOS activity of Chinese hamster
ovary cells was measured by the arginine
Activation of Constitutive Nitric-oxide Synthase Activity Is an
Early Signaling Event Induced by Ionizing Radiation*
§,
,, and**
Department of Radiation Oncology, Virginia
Commonwealth University, Richmond, Virginia 23298-0058 and the
¶ Division of Neonatology, Northwestern University,
Chicago, Illinois 60611-3008
citrulline conversion
assay. Irradiation stimulated a transient activation of NOS with
maximal activity at 5 min of post-irradiation. Western blot analysis
and genetic manipulation by overexpression of wild type or dominant negative NOS mutant identify the radiation-induced isoform as NOS-1.
Further evidence that NOS-1 is activated by radiation was the
demonstration of radiation-induced cGMP formation in cells transiently
transfected with the NO-dependent soluble guanylate cyclase. Protein Tyr nitration, a footprint of peroxynitrite formation, followed radiation exposure and was inhibited by expression of a
dominant negative NOS-1 mutant. Radiation-induced ERK1/2 kinase activity, a cytoprotective response to radiation, was also blocked by
inhibiting NOS activity. These experiments establish
NO-dependent signal transduction pathways as being
radioresponsive. Given the lipophilic and relatively stable properties
of NO, these results also suggest a possible mechanism by which
ionization events in one cell may activate signaling processes in
adjacent cells.
*
This work was supported in part by United States Public
Health Service Grants CA65896, CA72955, and 5T32DK07150, developmental funds from the Massey Cancer Center, and a generous gift from Tanya Gordon.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Supported by United States Public Health Service Grants
HL60190 and HD398110.
**
To whom correspondence should be addressed. Tel.: 804-628-0857;
Fax: 804-828-6042; E-mail: rmikkels@vcu.edu.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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