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The PDZ-LIM protein CLP36 is required for actin stress fiber formation and focal adhesion assembly in BeWo cells
Authors:Tamura Naoaki  Ohno Koji  Katayama Taiichi  Kanayama Naohiro  Sato Kohji
Affiliation:a Department of Anatomy, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashiku, Hamamatsu, 431-3192 Shizuoka, Japan
b Department of Obstetrics and Gynaecology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashiku, Hamamatsu, 431-3192 Shizuoka, Japan
Abstract:
CLP36 belongs to the ALP subfamily of PDZ-LIM proteins and has a PDZ domain at its N-terminal and a LIM domain at its C-terminal. It has been shown that CLP36 is localized to stress fibers through interaction with α-actinin, but its function is still unclear. To investigate the role of CLP36 in stress fibers, we suppressed CLP36 expression in BeWo cells by RNAi and examined the phenotypic changes. CLP36-knockdown resulted in cell spreading and the loss of stress fibers and focal adhesions. These changes were reversed by addition of exogenous CLP36, but not by addition of mutant forms of CLP36 that lacked the PDZ or LIM domain. These findings indicate that CLP36 plays a critical role in stress fiber formation and the assembly of focal adhesions in BeWo cells. In addition, the PDZ and LIM domains are both essential for CLP36 to function.
Keywords:CLP36   Pdlim1   Adaptor protein   RNAi   F-actin
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