原创力文档a mutant of swap-70, a phosphatidylinositoltrisphosphate binding protein, transforms mouse embryo fibroblasts, which is inhibited by sanguinarine突变的交换- 70,phosphatidylinositoltrisphosphate结合蛋白,将小鼠胚胎成纤维细胞,抑制血根碱.pdfVIP
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a mutant of swap-70, a phosphatidylinositoltrisphosphate binding protein, transforms mouse embryo fibroblasts, which is inhibited by sanguinarine突变的交换- 70,phosphatidylinositoltrisphosphate结合蛋白,将小鼠胚胎成纤维细胞,抑制血根碱
A Mutant of SWAP-70, a
Phosphatidylinositoltrisphosphate Binding Protein,
Transforms Mouse Embryo Fibroblasts, Which Is
Inhibited by Sanguinarine
Yasuhisa Fukui1,2*, Sayoko Ihara3
1 Laboratory of Signal Transduction, Hoshi University, Tokyo, Japan, 2 Institute of Cellular and Systems Medicine, National Health Research Institutes, Zhunan, Taiwan,
Republic of China, 3 Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo, Japan
Abstract
SWAP-70, a phosphatidylinositol trisphosphate (PtdIns(3,4,5)P ) binding protein, has been suggested to be involved in
3
transformation of mouse embryo fibroblasts (MEFs) as well as membrane ruffling after growth factor stimulation of the cells.
A mutant, SWAP-70-374, was found to be able to bind to F-actin in vitro, whereas wild-type SWAP-70 failed to do so. This
mutant was present at the plasma membrane without any stimulation while the wild-type protein was present only in the
cytosol unless cells were stimulated with EGF. Expression of this mutant in MEFs resulted in morphologic transformation,
fast growth, and loss of contact inhibition, suggesting that SWAP-70 with this mutation can transform the cells. ERK1/2 was
activated in SWAP-70-374-transformed cells. Use of MEK inhibitors revealed that the ERK1/2 pathway does not affect the cell
growth of MEFs but is responsible for loss of contact inhibition. To investigate the function of SWAP-70 further, drugs that
can inhibit SWAP-70-dependent cell responses were screened. Among various drugs, sanguinarine was found to inhibit
transformation of MEFs by SWAP-70-374. This drug was able to inhibit SWAP-70-mediated membrane ruffling as well,
suggesting that its effect was closely related to the SWAP-70 signaling pathway. These results suggest that SWAP-70-374
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