Association of Cell Adhesion Kinase bata (CAKbata/PYK2) with Cytoskeleton

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概要

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• Cell adhesion kinase bata (CAKbata/PYK2), which we identified by cDNA cloning, is the second protein-tyrosine kinase of the focal adhesion kinase subfamily and and has large N- and C-terminal domains in addition to the central kinase domain,In this paper, we report that CAKbata is present in association with both microtubules and microfilaments. The intracellular localization of CAKbata was studied in epithelial cells (A-431, Caco-2, and MDCK) and in nonepithelial cells (PC-12h and WFB), The immunocytochemical staining of epithelial cells revealed that CAKbata was mainly found at the cell-to-cell borders and at the perinuclear regions, although focal adhesions were also stained to some extent, The cell-to-cell immunostained in fine dotted lines and a significant portion of CAKbata was found in association with the microtubules, Moreever, an association of CAKbata with microfilaments was found in the cytoplasm, Among WFB cells only a limited number were positive for the CAKbata immunostaining at the focal contacts, although the protein was present in the tips of lamelipodia and microspikes in many cells, Furthermore, CAKbata was concentrated at the edge of ruffling membranes, When A431 cells were stimulated with epidermal growth factor, CAKbata rapidly moved to the edges of the ruffling membranes from the sites of cell-cell contacts where the protein had been present before the stimulation, Anti-actin or ati-alpha-tubulin antibodies coimmunoprecipitated CAKbata from a cytoskeleton-rich fraction of WFB cells. Moreover,the amount of CAKbata increased when the immunoprecipitation was carried out with the fraction prepared from the cells after stabilizing the tubular structure of the cytoskeleton.Imunoelectron microscopy showed that CAKbata was present along the microfilaments. In the WFB cells transfected with recombinant adenovirus expressing CAKbata, the protein was found at the focal adhesions of many cells and at the perinuclear region of all cells. Thus, CAKbata may associate with both microfilaments and microtubules, and play important roles for cellular movements such as membrane ruffling, cellular attachments, and cytokinesis

• 札幌医科大学の論文

著者

• Sasaki Hiroko

  Department of Obstetrics and Gynecology, Kobe University Graduate School of Medicine

• AOTO Hiroshi

  Department of Applied Chemistry, Faculty of Engineering, Osaka Institute of Technology

• Aoto Hiroshi

  Department Of Biochemistry Cancer Research Institute Sapporo Medical University

• Aoto Hiroshi

  Department Of Applied Chemistry Faculty Of Engineering Osaka Institute Of Technology

• Aoto Hiroshi

  Departments Of Biochemistry Cancer Research Inatitute Sapporo Medical University School Of Medicine

• MITAKA Toshihiro

  Department of Pathology, Cancer Research Institute, Sapporo Medical University School of Medicine

• Sasaki Hiroko

  Department Of Biochemistry Cancer Research Institute Sapporo Medical University School Of Medicine

• Mitaka Toshihiro

  Department Of Molecular Pathophysiology Cancer Research Institute Sapporo Medical University School

• Matsuno Takashi

  Department Of Surgery Sapporo Medical University

• Sasaki Hiroko

  Departments Of Biochemistry Cancer Research Inatitute Sapporo Medical University School Of Medicine

• Mitaka Toshihiro

  First Department Of Surgery Sapporo Medical University School Of Medicine

• Mitaka Toshihiro

  Department Of Pathology Cancer Research Institute Sapporo Medical University School Of Medicine

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