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

Boczonadi, Veronika; McInroy, Lorna; Maatta, Arto

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.
• Public URL: https://durham-repository.worktribe.com/output/1570824