Abstract
Reactive oxygen species (ROS) are well known to be involved in oncogene-mediated cellular transformation. However, the regulatory mechanisms underlying ROS generation in oncogene-transformed cells are unclear. In the present study, we found that oncogenic K-Ras induces ROS generation through activation of NADPH oxidase 1 (NOX1), which is a critical regulator for the K-Ras-induced cellular transformation. NOX1 was activated by K-Ras-dependent translocation of p47phox, a subunit of NOX1 to plasma membrane. Of note, PKCδ, when it was activated by PDPK1, directly bound to the SH3-N domain of p47phox and catalyzed the phosphorylation on Ser348 and Ser473 residues of p47phox C-terminal in a K-Ras-dependent manner, finally leading to its membrane translocation. Notably, oncogenic K-Ras activated all MAPKs (JNK, ERK and p38); however, only p38 was involved in p47phox-NOX1-dependent ROS generation and consequent transformation. Importantly, K-Ras-induced activation of p38 led to an activation of PDPK1, which then signals through PKCδ, p47phox and NOX1. In agreement with the mechanism, inhibition of p38, PDPK1, PKCδ, p47phox or NOX1 effectively blocked K-Ras-induced ROS generation, anchorage-independent colony formation and tumor formation. Taken together, our findings demonstrated that oncogenic K-Ras activates the signaling cascade p38/PDPK1/PKCδ/p47phox/NOX1 for ROS generation and consequent malignant cellular transformation.
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Abbreviations
- PKC:
-
protein kinase C
- PDPK1:
-
3-phosphoinositide-dependent protein kinase-1
- MAPK:
-
mitogen-activated protein kinase
- NADPH:
-
nicotinamide adenine dinucleotide phosphate-oxidase
- DN:
-
dominant-negative mutant form
- ROS:
-
reactive oxygen species
- GST:
-
glutathione S-transferases
- siRNA:
-
small interfering RNA
- WT:
-
wild type
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Acknowledgements
This work was supported by the National Research Foundation (NRF) and Ministry of Science, ICT and Future Planning, Korean government, through its National Nuclear Technology Program (2012M2A2A7035878 and 2012M2B2B1055639).
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Park, MT., Kim, MJ., Suh, Y. et al. Novel signaling axis for ROS generation during K-Ras-induced cellular transformation. Cell Death Differ 21, 1185–1197 (2014). https://doi.org/10.1038/cdd.2014.34
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DOI: https://doi.org/10.1038/cdd.2014.34
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