ABSTRACT
AKT phosphorylates components of the intrinsic cell survival machinery and promotes survival to various stimuli. In the present study, we identified CDC-like kinase 2 (CLK2) as a new substrate of AKT activation and elucidated its role in cell survival to ionizing radiation. AKT directly binds to and phosphorylates CLK2 on serine 34 and threonine 127, in vitro and in vivo. CLK2 phosphorylation was detected in HeLa cells overexpressing active AKT. In addition, we demonstrated that ionizing radiation induces CLK2 phosphorylation via AKT activation. In contrast, the suppression of endogenous AKT expression by siRNA inhibited CLK2 phosphorylation in response to 2 gray of γ-ray or insulin. Furthermore, we examined the effect of CLK2 on the survival of irradiated CCD-18Lu cells overexpressing Myc-CLK2. CLK2 overexpression significantly increased cell growth and inhibited cell death induced by 2 gray. The role of CLK2 in cell survival to ionizing radiation was dependent on the phosphorylation of serine 34 and threonine 127. Our results suggest that AKT activation controls cell survival to ionizing radiation by phosphorylating CLK2, revealing an important regulatory mechanism required for promoting cell survival.
Subject(s)
Gamma Rays/adverse effects , Protein Serine-Threonine Kinases/metabolism , Protein-Tyrosine Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Cell Death/drug effects , Cell Death/genetics , Cell Death/radiation effects , Cell Survival/drug effects , Cell Survival/genetics , Cell Survival/radiation effects , Enzyme Activation/drug effects , Enzyme Activation/genetics , Enzyme Activation/radiation effects , Gene Silencing , HeLa Cells , Humans , Hypoglycemic Agents/pharmacology , Insulin/pharmacology , Phosphorylation/drug effects , Phosphorylation/genetics , Phosphorylation/radiation effects , Protein Serine-Threonine Kinases/genetics , Protein-Tyrosine Kinases/genetics , Proto-Oncogene Proteins c-akt/genetics , RNA, Small InterferingABSTRACT
1-Bromopropane (1-BP) has recently become known as an alternative cleaning material with less damage to the ozone layer. However, its toxicity is not fully evaluated. This study was designed to investigate the repeated inhalation toxicity of 1-BP on the nervous systems in Sprague-Dawley rats. The experiment was done by repeated exposure of the rats to 0, 200, 500, and 1250 ppm for 6 h per day, 5 days a week, for 13 weeks, respectively. Morphologic studies were done for the central nervous system, sacral and peroneal nerves. The serial sections of the brain and spinal cord of 1-BP inhalation groups revealed no pathological features either in the gray or white matter. The nerve fiber teasing, light and electron microscopic studies of the sacral and peroneal nerve fibers showed no significant difference between 1-BP inhalation groups and the control group. From these results, it is concluded that the nervous system is histologically resistant to the repeated inhalation of 1-BP up to 1250 ppm for 13 weeks. Experiments with higher concentrations of 1-BP and the functional studies are necessary to clarify the 1-BP toxicity.
