Rescue of atypical protein kinase C in epithelia by the cytoskeleton and Hsp70 family chaperones

Anastasia Mashukova, Andrea S. Oriolo, Flavia A. Wald, M. Llanos Casanova, Cornelia Kröger, Thomas M. Magin, M. Bishr Omary, Pedro J.I. Salas

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

Atypical PKC (PKCι) is a key organizer of cellular asymmetry. Sequential extractions of intestinal cells showed a pool of enzymatically active PKCι and the chaperone Hsp70.1 attached to the apical cytoskeleton. Pull-down experiments using purified and recombinant proteins showed a complex of Hsp70 and atypical PKC on filamentous keratins. Transgenic animals overexpressing keratin 8 displayed delocalization of Hsp70 and atypical PKC. Two different keratin-null mouse models, as well as keratin-8 knockdown cells in tissue culture, also showed redistribution of Hsp70 and a sharp decrease in the active form of atypical PKC, which was also reduced by Hsp70 knockdown. An in-vitro turn motif rephosphorylation assay indicated that PKCι is dephosphorylated by prolonged activity. The Tritonsoluble fraction could rephosphorylate PKCι only when supplemented with the cytoskeletal pellet or filamentous highly purified keratins, a function abolished by immunodepletion of Hsp70 but rescued by recombinant Hsp70. We conclude that both filamentous keratins and Hsp70 are required for the rescue rephosphorylation of mature atypical PKC, regulating the subcellular distribution and steady-state levels of active PKCι.

Original languageEnglish (US)
Pages (from-to)2491-2503
Number of pages13
JournalJournal of Cell Science
Volume122
Issue number14
DOIs
StatePublished - Jul 15 2009

Keywords

  • Apical kinases
  • Epithelial polarity
  • Keratins
  • PKCλ
  • Protein chaperones

ASJC Scopus subject areas

  • Cell Biology

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    Mashukova, A., Oriolo, A. S., Wald, F. A., Casanova, M. L., Kröger, C., Magin, T. M., Omary, M. B., & Salas, P. J. I. (2009). Rescue of atypical protein kinase C in epithelia by the cytoskeleton and Hsp70 family chaperones. Journal of Cell Science, 122(14), 2491-2503. https://doi.org/10.1242/jcs.046979