Proteomic profiling of human keratinocytes undergoing UVB-induced alternative differentiation reveals tripartite motif protein 29 as a survival factor

Véronique Vallery, Nathalie Belot, Marc Dieu, Edouard Delaive, Noëlle Ninane, Catherine Demazy, Martine Raes, Michel Salmon, Yves Poumay, Florence Debacq-Chainiaux, Olivier Toussaint

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Abstract

Background: Repeated exposures to UVB of human keratinocytes lacking functional p16INK-4a and able to differentiate induce an alternative state of differentiation rather than stress-induced premature senescence. Methodology/Principal Findings: A 2D-DIGE proteomic profiling of this alternative state of differentiation was performed herein at various times after the exposures to UVB. Sixty-nine differentially abundant protein species were identified by mass spectrometry, many of which are involved in keratinocyte differentiation and survival. Among these protein species was TRIpartite Motif Protein 29 (TRIM29). Increased abundance of TRIM29 following UVB exposures was validated by Western blot using specific antibody and was also further analysed by immunochemistry and by RT-PCR. TRIM29 was found very abundant in keratinocytes and reconstructed epidermis. Knocking down the expression of TRIM29 by short-hairpin RNA interference decreased the viability of keratinocytes after UVB exposure. The abundance of involucrin mRNA, a marker of late differentiation, increased concomitantly. In TRIM29-knocked down reconstructed epidermis, the presence of picnotic cells revealed cell injury. Increased abundance of TRIM29 was also observed upon exposure to DNA damaging agents and PKC activation. The UVB-induced increase of TRIM29 abundance was dependent on a PKC signaling pathway, likely PKCd. Conclusions/Significance: These findings suggest that TRIM29 allows keratinocytes to enter a protective alternative differentiation process rather than die massively after stress. © 2010 Bertrand-Vallery et al.

Original languageEnglish
Article numbere10462
JournalPLoS ONE
Volume5
Issue number5
DOIs
Publication statusPublished - 14 Sep 2010

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