title: Binding of the extreme carboxyl-terminus of PAK-interacting exchange factor β (βPIX) to myosin 18A (MYO18A) is required for epithelial cell migration.
abstract: The PAK2/βPIX/GIT1 (p21-activated kinase 2/PAK-interacting exchange factor-β/G protein-coupled receptor kinase-interactor 1) complex has been shown to distribute to both membrane ruffles and focal adhesions of cells, where it plays an important role in regulating focal adhesion turnover. However, the detailed mechanism underlying this regulation is largely unknown. We previously reported that MYO18Aα interacts via its carboxyl terminus with the PAK2/βPIX/GIT1 complex through direct binding to βPIX, and that knockdown of MYO18Aα in epithelial cells causes accumulation of the complex in focal adhesions and decreased cell migration ability (Hsu et al., 2010). The current study characterized the detailed MYO18Aα-βPIX interaction mechanism and the biological significance of this interaction. We found that deletion of the carboxyl-terminal globular domain of MYO18Aα profoundly altered the cellular localization of βPIX and inhibited cell migration. βPIX interacts through its most carboxyl-terminus, PAWDETNL (639-646), with MYO18Aα and partially colocalized with MYO18Aα in membrane ruffles of cells, whereas βPIX(1-638), a mutant with deletion of PAWDETNL, accumulated in focal adhesions. Both focal adhesion numbers and area in βPIX(1-638)-expressing cells were greater than those in cells expressing wild-type βPIX(FL). Further experiments using deletion mutants of MYO18A and βPIX showed that disruption of MYO18A-βPIX interaction not only impaired cell motility but also decreased Rac1 activity. Collectively, our data unravel the interaction regions between MYO18A and βPIX and provide evidence for the critical role of this interaction in regulating cellular localization of βPIX, Rac1 activity, and adhesion and migration in epithelial cells.
authors.affiliation: Department of Cell and Molecular Biology, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan, ROC.
authors.forename: Rae-Mann
authors.initials: RM
authors.lastname: Hsu
authors.affiliation: Molecular Medicine Research Center, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan, ROC.
authors.forename: Ya-Ju
authors.initials: YJ
authors.lastname: Hsieh
authors.affiliation: Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan, ROC.
authors.forename: Tsung-Han
authors.initials: TH
authors.lastname: Yang
authors.affiliation: Department of Cell and Molecular Biology, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan, ROC.
authors.forename: Yi-Chien
authors.initials: YC
authors.lastname: Chiang
authors.affiliation: Department of Cell and Molecular Biology, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan, ROC.
authors.forename: Chih-Yen
authors.initials: CY
authors.lastname: Kan
authors.affiliation: Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan, ROC.
authors.forename: Yu-Tsuen
authors.initials: YT
authors.lastname: Lin
authors.affiliation: Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan, ROC.
authors.forename: Jeng-Ting
authors.initials: JT
authors.lastname: Chen
authors.affiliation: Department of Cell and Molecular Biology, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan, ROC; Molecular Medicine Research Center, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan, ROC; Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan, ROC. Electronic address: yusong@mail.cgu.edu.tw.
authors.forename: Jau-Song
authors.initials: JS
authors.lastname: Yu
keywords: Cell migration; Focal adhesion; GIT1; MYO18A; PAK2; βPIX