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Ultraviolet B irradiation and activation of protein kinase D in primary mouse epidermal keratinocytes

Oncogene volume 30, pages 1586–1596 (2011)Cite this article

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Abstract

Our previous studies demonstrated that protein kinase D (PKD), a serine/threonine kinase implicated in various cell processes, is upregulated in basal cell carcinoma (BCC), supporting a possible tumorigenic role for PKD in skin. As the greatest risk factor for BCC is sun exposure, the ability of ultraviolet B (UVB) irradiation to activate PKD in primary mouse keratinocytes was investigated. Using western analysis with two autophosphorylation-specific antibodies, we show for the first time that UVB activated PKD in a time- and dose-dependent manner. UVB-induced PKD activation was verified using an in vitro kinase assay. Furthermore, activation was reduced by antioxidant pretreatment, suggesting a link with oxidative stress. UVB-induced PKD activation was mediated primarily by Src family tyrosine kinases rather than protein kinase C (PKC), and in fact, UVB did not alter PKC-mediated transphosphorylation. UVB induced apoptosis dose dependently, and this death could be prevented by overexpression of wild-type PKD, but not mutant PKD or the empty adenovirus. Indeed, a mutant that cannot be phosphorylated by Src kinases exacerbated UVB-elicited apoptosis. Thus, our data indicate that UVB irradiation of keratinocytes induces Src-mediated activation of PKD, which protects cells from UVB-stimulated apoptosis, providing a possible explanation for the observed upregulation of PKD in BCC.

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Acknowledgements

This project was supported by a Merit Award from the Veterans’ Administration and grants from the National Institutes of Health/National Institute of Arthritis, Musculoskeletal and Skin Diseases #AR45212 and AR57321 (WBB). We thank Dr Alex Toker for generously providing the various PKD constructs as well as for his useful discussions, and Dr Bert Vogelstein for his kind gift of the AdEasy adenovirus system. We appreciate the expert technical assistance of Mr Peter Parker and Ms Mariya Wilson. This work was submitted in partial fulfilment of the requirements for a doctoral degree from the Medical College of Georgia (SNA).

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  1. B A Shapiro

    Present address: 4Current address: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, VA, USA.,

Authors and Affiliations

  1. Institute of Molecular Medicine and Genetics, Medical College of Georgia, Augusta, GA, USA

    S N Arun, B A Shapiro & W B Bollag

  2. Charlie Norwood VA Medical Center, One Freedom Way, Augusta, GA, USA

    I Kaddour-Djebbar & W B Bollag

  3. Departments of Physiology and Medicine (Dermatology), Medical College of Georgia, Augusta, GA, USA

    W B Bollag

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  1. S N Arun
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Correspondence to W B Bollag.

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Arun, S., Kaddour-Djebbar, I., Shapiro, B. et al. Ultraviolet B irradiation and activation of protein kinase D in primary mouse epidermal keratinocytes. Oncogene 30, 1586–1596 (2011). https://doi.org/10.1038/onc.2010.540

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