TY - JOUR
T1 - Spatial regulation of MCAK promotes cell polarization and focal adhesion turnover to drive robust cell migration
AU - Zong, Hailing
AU - Hazelbaker, Mark
AU - Moe, Christina
AU - Ems-McClung, Stephanie C.
AU - Hu, Ke
AU - Walczak, Claire E.
N1 - Funding Information:
This work was supported by National Institutes of Health grant R35GM122482 to C.E.W. We thank Anne Straube for paxillin-GFP– expressing RPE-1 cells, Yeajin Kim and Weini Wang for help in optimizing centrosome orientation experiments, Ben Walker for assistance with the centrosome positioning algorithm, and Sanjay Shrestha for technical assistance in cell culture. We also thank Jim Powers for help with imaging and Sid Shaw for thoughtful discussions of the data and analysis. The Indiana University–Light Microscopy Imaging Center is supported in part by the Office of the Vice Provost for Research.
Publisher Copyright:
© 2021 Zong et al.
PY - 2021/4/1
Y1 - 2021/4/1
N2 - The asymmetric distribution of microtubule (MT) dynamics in migrating cells is important for cell polarization, yet the underlying regulatory mechanisms remain underexplored. Here, we addressed this question by studying the role of the MT depolymerase, MCAK (mitotic centromere-associated kinesin), in the highly persistent migration of RPE-1 cells. MCAK knockdown leads to slowed migration and poor directional movement. Fixed and live cell imaging revealed that MCAK knockdown results in excessive membrane ruffling as well as defects in cell polarization and the maintenance of a major protrusive front. Additionally, loss of MCAK increases the lifetime of focal adhesions by decreasing their disassembly rate. These functions correlate with a spatial distribution of MCAK activity, wherein activity is higher in the trailing edge of cells compared with the leading edge. Overexpression of Rac1 has a dominant effect over MCAK activity, placing it downstream of or in a parallel pathway to MCAK function in migration. Together, our data support a model in which the polarized distribution of MCAK activity and subsequent differential regulation of MT dynamics contribute to cell polarity, centrosome positioning, and focal adhesion dynamics, which all help facilitate robust directional migration.
AB - The asymmetric distribution of microtubule (MT) dynamics in migrating cells is important for cell polarization, yet the underlying regulatory mechanisms remain underexplored. Here, we addressed this question by studying the role of the MT depolymerase, MCAK (mitotic centromere-associated kinesin), in the highly persistent migration of RPE-1 cells. MCAK knockdown leads to slowed migration and poor directional movement. Fixed and live cell imaging revealed that MCAK knockdown results in excessive membrane ruffling as well as defects in cell polarization and the maintenance of a major protrusive front. Additionally, loss of MCAK increases the lifetime of focal adhesions by decreasing their disassembly rate. These functions correlate with a spatial distribution of MCAK activity, wherein activity is higher in the trailing edge of cells compared with the leading edge. Overexpression of Rac1 has a dominant effect over MCAK activity, placing it downstream of or in a parallel pathway to MCAK function in migration. Together, our data support a model in which the polarized distribution of MCAK activity and subsequent differential regulation of MT dynamics contribute to cell polarity, centrosome positioning, and focal adhesion dynamics, which all help facilitate robust directional migration.
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U2 - 10.1091/MBC.E20-05-0301
DO - 10.1091/MBC.E20-05-0301
M3 - Article
C2 - 33566676
AN - SCOPUS:85103800333
VL - 32
SP - 590
EP - 604
JO - Molecular Biology of the Cell
JF - Molecular Biology of the Cell
SN - 1059-1524
IS - 7
ER -