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Cytolinker Cross-talk: Periplakin N-terminus Interacts with Plectin to Regulate Keratin Organisation and Epithelial Migration

Boczonadi, Veronika; McInroy, Lorna; Maatta, Arto

Authors

Veronika Boczonadi

Lorna McInroy



Abstract

Periplakin is a cytoskeletal linker protein that participates in the assembly of epidermal cell cornified envelope and regulates keratin organisation in simple epithelial cells. We have generated a stably transfected MCF-7 subclone expressing HA-tagged periplakin N-terminus to identify molecular interactions of periplakin. Co-immunoprecipitation with anti-HA antibodies and mass spectrometry identified a 500-kDa periplakin-interacting protein as plectin, another plakin family member. Plectin–periplakin interaction was confirmed by immunoblotting of complexes immunoprecipitated by either anti-HA or anti-plectin antibodies. Transient transfections of periplakin deletion constructs indicated that first 133 amino acid residues of the N-terminus are sufficient for co-localisation with plectin at MCF-7 cell borders. Immunofluorescence analysis demonstrated that periplakin and plectin isoforms 1, 1f and 1k co-localise at cell borders of MCF-7 epithelia and that plectin-1f and 1k co-localise with periplakin in suprabasal epidermis. Ablation of plectin by siRNA in HaCaT keratinocytes resulted in aggregation of periplakin to small clusters. Scratch-wounded MCF-7 epithelia expressing periplakin N-terminus showed accelerated keratin re-organisation that was inhibited by siRNA knock-down of plectin. Finally, ablation of either periplakin or plectin, or both proteins simultaneously, impaired migration of MCF-7 epithelial sheets. Thus, we have identified a novel functional co-localisation between two plakin cytolinker proteins.

Citation

Boczonadi, V., McInroy, L., & Maatta, A. (2007). Cytolinker Cross-talk: Periplakin N-terminus Interacts with Plectin to Regulate Keratin Organisation and Epithelial Migration. Experimental Cell Research, 313(16), 3579-3591. https://doi.org/10.1016/j.yexcr.2007.07.005

Journal Article Type Article
Publication Date Oct 1, 2007
Deposit Date Sep 21, 2007
Journal Experimental Cell Research
Print ISSN 0014-4827
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 313
Issue 16
Pages 3579-3591
DOI https://doi.org/10.1016/j.yexcr.2007.07.005
Keywords Plectin, Periplakin, Epithelial migration, Intermediate filaments.