期刊
FASEB JOURNAL
卷 37, 期 9, 页码 -出版社
WILEY
DOI: 10.1096/fj.202201992R
关键词
actin dynamics; high-speed atomic force microscopy; mechano-response; Rho GTPase-activating protein; Yes-associated protein
This study utilized high-speed atomic force microscopy (HS-AFM) and other techniques to examine the structure and dynamics of intracellular actin architecture in normal and YAP-knockout cells. The results showed that YAP influences the density, dynamics, and elasticity of actin filaments in cells, as well as the migration and activity of ARHGAP18, leading to changes in cell architecture and motility.
Yes-associated protein (YAP) is a transcriptional co-activator that controls the transcription of target genes and modulates the structures of various cytoskeletal architecture as mechanical responses. Although it has been known that YAP regulates actin-regulatory proteins, the detailed molecular mechanism of how they control and coordinate intracellular actin architecture remains elusive. Herein, we aimed to examine the structure and dynamics of intracellular actin architecture from molecular to cellular scales in normal and YAP-knockout (YAP-KO) cells utilizing high-speed atomic force microscopy (HS-AFM) for live-cell imaging and other microscope-based mechanical manipulation and measurement techniques. YAP-KO Madin-Darby canine kidney cells had a higher density and turnover of actin filaments in the cell cortex and a higher elastic modulus. Laser aberration assay demonstrated that YAP-KO cells were more resistant to damage than normal cells. We also found that Rho GTPase-activating protein 18 (ARHGAP18), a downstream factor of YAP, translocated from the cortex to the edge of sparsely cultured YAP-KO cells. It resulted in high RhoA activity and promotion of actin polymerization in the cell cortex and their reductions at the edge. HS-AFM imaging of live cell edge and a cell-migration assay demonstrated lower membrane dynamics and motility of YAP-KO cells than those of normal cells, suggesting lower actin dynamics at the edge. Together, these results demonstrate that a YAP-dependent pathway changes the intracellular distribution of RhoGAP and modulates actin dynamics in different parts of the cell, providing a mechanistic insight into how a mechano-sensitive transcription cofactor regulates multiple intracellular actin architecture and coordinates mechano-responses.
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