A coordinated phosphorylation by Lats and CK1 regulates YAP stability through SCF(beta-TRCP)

Department of Pharmacology and Moores Cancer Center, University of California at San Diego, La Jolla, California 92093-0815, USA.
Genes & development (Impact Factor: 10.8). 01/2010; 24(1):72-85. DOI: 10.1101/gad.1843810
Source: PubMed


The Yes-associated protein (YAP) transcription coactivator is a key regulator of organ size and a candidate human oncogene. YAP is inhibited by the Hippo pathway kinase cascade, at least in part via phosphorylation of Ser 127, which results in YAP 14-3-3 binding and cytoplasmic retention. Here we report that YAP is phosphorylated by Lats on all of the five consensus HXRXXS motifs. Phosphorylation of Ser 381 in one of them primes YAP for subsequent phosphorylation by CK1delta/epsilon in a phosphodegron. The phosphorylated phosphodegron then recruits the SCF(beta-TRCP) E3 ubiquitin ligase, which catalyzes YAP ubiquitination, ultimately leading to YAP degradation. The phosphodegron-mediated degradation and the Ser 127 phosphorylation-dependent translocation coordinately suppress YAP oncogenic activity. Our study identified CK1delta/epsilon as new regulators of YAP and uncovered an intricate mechanism of YAP regulation by the Hippo pathway via both S127 phosphorylation-mediated spatial regulation (nuclear-cytoplasmic shuttling) and the phosphodegron-mediated temporal regulation (degradation).

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    • "There is no change in the amount of EDTB, YAP, AMOT, or AMOTL2 in cells, regardless of confluence (Figure 5A). This contrasts with previous reports, in which levels of the YAP were found to be either up or down under confluent conditions (Zhao et al., 2007, 2010). This may relate to cell-type differences, as experiments in these studies were performed in fibroblasts. "
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    ABSTRACT: Contact-mediated inhibition of cell proliferation is an essential part of organ growth control; the transcription coactivator Yes-associated protein (YAP) plays a pivotal role in this process. In addition to phosphorylation-dependent regulation of YAP, the integral membrane protein Angiomotin (AMOT) and AMOT family members control YAP through direct binding. Here we report that regulation of YAP activity occurs at the endosomal membrane through a dynamic interaction of AMOT with an endosomal integral membrane protein endotubin (EDTB). EDTB interacts with both AMOT and occludin and preferentially associates with occludin in confluent cells but with AMOT family members in subconfluent cells. EDTB competes with YAP for binding to AMOT proteins in subconfluent cells. Over-expression of the cytoplasmic domain or full-length EDTB induces translocation of YAP to the nucleus, an overgrowth phenotype and growth in soft agar. This increase in proliferation is dependent upon YAP activity and is complemented by overexpression of p130-AMOT. Furthermore, overexpression of EDTB inhibits the AMOT:YAP interaction. EDTB and AMOT have a greater association in subconfluent cells compared with confluent cells, and this association is regulated at the endosomal membrane. These data provide a link between the trafficking of tight junction proteins through endosomes and contact-inhibition-regulated cell growth. © 2015 by The American Society for Cell Biology.
    Molecular biology of the cell 05/2015; 26(14). DOI:10.1091/mbc.E15-04-0224 · 4.47 Impact Factor
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    • "JEM Vol. 211, No. 11 Ar ticle 2255 of YAP in Tsc2-null cells, because the amount of Ser112- phosphorylated YAP, a site of the Lats1/2 kinases (Zhao et al., 2010), paralleled the increase in total YAP protein in the mutant cells (Fig. 5 a). Tsc2 deletion is known to provide a proliferative advantage over wild-type controls in serum starvation conditions (Zhang et al., 2003). "
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    ABSTRACT: Genetic studies have shown that the tuberous sclerosis complex (TSC) 1-TSC2-mammalian target of Rapamycin (mTOR) and the Hippo-Yes-associated protein 1 (YAP) pathways are master regulators of organ size, which are often involved in tumorigenesis. The crosstalk between these signal transduction pathways in coordinating environmental cues, such as nutritional status and mechanical constraints, is crucial for tissue growth. Whether and how mTOR regulates YAP remains elusive. Here we describe a novel mouse model of TSC which develops renal mesenchymal lesions recapitulating human perivascular epithelioid cell tumors (PEComas) from patients with TSC. We identify that YAP is up-regulated by mTOR in mouse and human PEComas. YAP inhibition blunts abnormal proliferation and induces apoptosis of TSC1-TSC2-deficient cells, both in culture and in mosaic Tsc1 mutant mice. We further delineate that YAP accumulation in TSC1/TSC2-deficient cells is due to impaired degradation of the protein by the autophagosome/lysosome system. Thus, the regulation of YAP by mTOR and autophagy is a novel mechanism of growth control, matching YAP activity with nutrient availability under growth-permissive conditions. YAP may serve as a potential therapeutic target for TSC and other diseases with dysregulated mTOR activity.
    Journal of Experimental Medicine 10/2014; 211(11). DOI:10.1084/jem.20140341 · 12.52 Impact Factor
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    • "Activation of the Hippo pathway leads to phosphorylation of LATS1/2 by MST1/2. LATS1/2 in turn phosphorylate YAP on multiple sites, including S127, which promotes cytoplasmic retention of YAP (Zhao et al., 2007; Hao et al., 2008; Lei et al., 2008), and S381 (S397 in human YAP1), which promotes YAP turnover (Zhao et al., 2010b). Expression of ST decreased YAP S127 phosphorylation in BJ fibroblasts and in HMECs (Figure 2A). "
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    ABSTRACT: Primary human cells can be transformed into tumor cells by a defined set of genetic alterations including telomerase, oncogenic Ras(V12), and the tumor suppressors p53 and pRb. SV40 small T (ST) is required for anchorage-independent growth in vitro and in vivo. Here, we identify the Hippo tumor suppressor pathway as a critical target of ST in cellular transformation. We report that ST uncouples YAP from the inhibitory activity of the Hippo pathway through PAK1-mediated inactivation of NF2. Membrane-tethered activated PAK is sufficient to bypass the requirement for ST in anchorage-independent growth. PAK acts via YAP to mediate the transforming effects of ST. Activation of endogenous YAP is required for ST-mediated transformation and is sufficient to bypass ST in anchorage-independent growth and xenograft tumor formation. Our findings uncover the Hippo tumor suppressor pathway as a final gatekeeper to transformation and tumorigenesis of primary cells.
    Cell Reports 07/2014; 8(3). DOI:10.1016/j.celrep.2014.06.062 · 8.36 Impact Factor
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