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Please use this identifier to cite or link to this item: http://hdl.handle.net/1974/1494

Title: A Novel Role for Calpain 4 in Podosome Assembly
Authors: Dowler, THOMAS

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Keywords: Calpain 4
Podosome assembly
Mouse embryonic fibroblast
Dorsal ruffles
Issue Date: 2008
Series/Report no.: Canadian theses
Abstract: Podosomes are adhesive and invasive structures which may play an important role in numerous physiological and pathological conditions including angiogenesis, atherosclerosis, and cancer metastasis. Recently, the cysteine protease m-calpain (m-Capn) has been shown to cleave cortactin, an integral component of the podosomal F-actin core, as well as various proteins found in the peripheral adhesive region leading to the disassembly of these dynamic structures. In this study, I investigated whether Capn plays a role in the formation of podosomes downstream of c-Src. I show that: 1) phorbol-12, 13-dibutyrate (PDBu) as well as c-Src-Y527F expression induces podosome formation in mouse embryonic fibroblasts; 2) PDBu- and constitutively active c-Src-induced podosome formation is inhibited by the knockout of the m- and µ-Capn small regulatory subunit Capn4 in mouse embryonic fibroblasts (Capn4-/-), but is partially restored by re-expression of Capn4; 3) Capn4 localizes to podosomes; and 4) Inhibition of m- and µ-Capn proteolytic activity by the cell permeable calpain inhibitors has little effect on the formation of podosomes downstream of active c-Src. I conclude that Capn4 may play a role in the assembly phase of podosomes independent of calpain proteolytic activity. Work done in collaboration to determine a possible mechanism of action for the role of Capn4 in podosome assembly indicates that a possible binding partner of Capn4, β-PIX, co-localizes with, and shows in vivo association with Capn4. Furthermore, β-PIX and Capn4 bind directly in vitro in the presence of Ca2+. We conclude that Capn4 plays a role in podosome assembly, and this role may be through direct interaction with β-PIX in a calcium-dependent manner.
Description: Thesis (Master, Biochemistry) -- Queen's University, 2008-09-26 16:16:00.768
URI: http://hdl.handle.net/1974/1494
Appears in Collections:Queen's Graduate Theses and Dissertations
Biochemistry Graduate Theses (July 2007 - Sept 2016)

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