Substrate rigidity-dependent positive feedback regulation between YAP and ROCK2

Abstract

Extracellular matrix (ECM) stiffness influences gene expression, leading to modulation of various cellular functions. While ROCK2 regulates actomyosin activity as well as cell migration and proliferation, expression of ROCK2 is increased in response to stiffening ECM. However, the mechanism underlying rigidity-dependent ROCK2 expression remains elusive. Here, we show that YAP, a mechanically regulated transcription coactivator, upregulates ROCK2 expression in an ECM rigidity-dependent manner. YAP interacted with the ROCK2 promoter region in an actomyosin activity-dependent manner. Knockdown of YAP decreased ROCK2 expression while activity of the ROCK2 promoter was upregulated by expressing constitutively active YAP. Furthermore, we found that ROCK2 expression promotes transcriptional activation by YAP. Our results reveal a novel positive feedback loop between YAP and ROCK2, which is modulated by ECM stiffness.

MMP24 as a Target of YAP Is a Potential Prognostic Factor in Cancer Patients

“…We have demonstrated that expression of the gene encoding Rho-associated coiled coilcontaining protein kinase (ROCK) 2, which induces the activation of myosin II, is increased in response to the rigid substrate in a YAP-TEAD dependent manner [9]. On the other hand, nuclear accumulation of YAP, in turn, depends on ROCK2 expression, as shown previously [10] and in Figure 3A. Therefore, we wondered whether ROCK2 played a role in the expression of MMP24 in cells cultured on the rigid substrate.…”

“…Similarly, the expression levels of two YAP target genes, CTGF and MMP7, were also elevated in cells grown on the rigid substrate ( Figure 1C and D). We have previously shown that rigid substrates promote YAPdependent transcription in MCF-7 cells [9,10]. We therefore examined whether YAP regulated MMP24 expression.…”

“…The nuclear translocation of YAP has been shown to be dependent upon actomyosin contractility [7][8][9][10]. We thus set out to investigate the role of actomyosin in the interaction between YAP and the MMP24 promoter.…”

“…The retrovirus vectors encoding small hairpin RNAs (shRNAs) against human YAP and human ROCK2, the YAP target sequences #1: 5 -GCCACCAAGCTAGATAAAGAA-3 , #2: 5 -GACA TCTTCTGGTCAGAGA-3 and ROCK2 target sequence: 5 -GGTTTATGCTATGAAGCTT-3 , were cloned into the pSuper retro puro vector (Oligoengine, Seattle, WA, USA) and pSuper retro hygro vector [10], respectively. Retroviral infection was performed as described previously [16].…”

“…Interaction of Yes-associated protein (YAP) and angiomotin (AMOT), which sequesters YAP in the cytoplasm, is attenuated by F-actin association to AMOT, resulting in the nuclear translocation of YAP [7,8]. Our previous study using MCF-7 cells has shown that an increase in actomyosin activity upregulates YAP-dependent gene expression [9,10]. Matrix metalloproteinases (MMPs) belong to a family of zinc-dependent endopeptidases and are categorized into two distinct classes: soluble MMPs and membrane-type (MT) MMPs [11].…”