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Oka T, Mazack V, Sudol M. Mst2 and Lats kinases regulate apoptotic function of Yes kinase-associated protein (YAP). J Biol Chem. 2008;283(41):27534-46

Laboratory of Signal Transduction and Proteomic Profiling, Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania 17822, USA.
Journal of Biological Chemistry (Impact Factor: 4.57). 10/2008; 283(41):27534-46. DOI: 10.1074/jbc.M804380200
Source: PubMed

ABSTRACT The Hippo pathway in Drosophila controls the size and shape of organs. In the fly, activation of this pathway conveys growth-inhibitory signals and promotes apoptosis in epithelial cells. We "reconstituted" the Hippo pathway in a human epithelial cell line and showed that, in contrast to flies, the activation of this pathway results in anti-apoptotic signals. We have shown that in human embryonic kidney (HEK) 293 cells, the complex formation between transcriptional co-activators YAPs (Yes kinase-associated proteins) and Lats kinases requires the intact WW domains of YAPs, as well as intact Pro-Pro-AA-Tyr (where AA is any amino acid) motifs in Lats kinases. These kinases cooperate with the upstream Mst2 kinase to phosphorylate YAPs at Ser-127. Overexpression of YAP2 in HEK293 cells promoted apoptosis, whereas the Mst2/Lats1-induced phosphorylation of YAP partially rescued the cells from apoptotic death. Apoptotic signaling of YAP2 was mediated via stabilization of p73, which formed a complex with YAP2. All components of the Hippo pathway that we studied were localized in the cytoplasm, with the exception of YAP, which also localized in the nucleus. The localization of YAP2 in the nucleus was negatively controlled by the Lats1 kinase. Our apoptotic "readout" of the Hippo pathway in embryonic kidney cells represents a useful experimental system for the identification of the putative upstream receptor, membrane protein, or extracellular factor that initiates an entire signaling cascade and ultimately controls the size of organs.

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    • "The STE20 like kinases MST1 and MST2 are core components of mammalian Hippo signaling pathway, which plays a fundamental role in organ size control and tumor suppression (Avruch et al., 2006; O'Neill and Kolch, 2005; O'Neill et al., 2004; Song et al., 2010). MST1/2 in conjunction with an adaptor protein Salvador phosphorylates and activates LATS1/2 kinases, which in turn associates with adaptor protein MOB1, to phosphorylate and prevent nuclear translocation of the downstream transcription coactivator Yes-associated protein (YAP), and thereby inhibit cell proliferation and promote apoptosis (Oka et al., 2008; Wu et al., 2003). MST1 and MST2 share high sequence homology with identical N-terminal kinase domains for catalysis and distinct C-terminal SARAH (Salvador, RASSF and Hpo homology) domains for homodimerization or hetero-interaction with Salvador and RASSF (Ras association domain family) proteins (Fig. 1A) (Pfeifer et al., 2010; Scheel and Hofmann, 2003). "
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    ABSTRACT: The STE20 kinases MST1 and MST2 are key players in mammalian Hippo pathway. The SARAH domains of MST1/2 act as a platform to mediate homodimerization and hetero-interaction with a range of adaptors including RASSFs and Salvador, which also possess SARAH domains. Here, we determined the crystal structure of human MST2 SARAH domain, which forms an antiparallel homodimeric coiled coil. Structural comparison indicates that SARAH domains of different proteins may utilize a shared dimerization module to form homodimer or heterodimer. Structure-guided mutational study identified specific interface residues critical for MST2 homodimerization. MST2 mutations disrupting its homodimerization also impaired its hetero-interaction with RAPL (also named RASSF5 and NORE1), which is mediated by their SARAH domains. Further biochemical and cellular assays indicated that SARAH domain-mediated homodimerization and hetero-interaction with RAPL are required for full activation of MST2 and therefore apoptotic functions in T cells.
    Journal of Structural Biology 01/2014; 186(1). DOI:10.1016/j.jsb.2014.01.008 · 3.23 Impact Factor
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    • "Cells were counted after trypsin detachment using a hemocytometer. MEFs were transfected with plasmids encoding Flag-Set7, Flag-Set7 H297G (a point mutation that abolishes Set7 methyltransferase activity) (Nishioka et al., 2002; Tao et al., 2011), Flag-Yap, Flag-Yap K494R , or Flag-Yap K497R (Addgene ''19045'') (Oka et al., 2008; Hata et al., 2012) constructs using Lipofectamine 2000 (Invitrogen) according to the manufacturer's descriptions. "
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    ABSTRACT: Methylation of nonhistone proteins is emerging as a regulatory mechanism to control protein function. Set7 (Setd7) is a SET-domain-containing lysine methyltransferase that methylates and alters function of a variety of proteins in vitro, but the in vivo relevance has not been established. We found that Set7 is a modifier of the Hippo pathway. Mice that lack Set7 have a larger progenitor compartment in the intestine, coinciding with increased expression of Yes-associated protein (Yap) target genes. Mechanistically, monomethylation of lysine 494 of Yap is critical for cytoplasmic retention. These results identify a methylation-dependent checkpoint in the Hippo pathway.
    Developmental Cell 07/2013; 26(2). DOI:10.1016/j.devcel.2013.05.025 · 10.37 Impact Factor
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    • "Genetic studies have established that Yki acts at the end of, and is inhibited by, the Hippo tumor suppressor pathway, which is highly conserved in mammals (Huang et al. 2005). YAP is phosphorylated and inhibited by the Lats1/2 kinases, which are key components of the Hippo pathway (Dong et al. 2007; Hao et al. 2007; Zhao et al. 2007; Oka et al. 2008; J Zhang et al. 2008). The phosphorylated YAP is retained in the cytoplasm, and therefore is inactive to stimulate gene expression. "
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    ABSTRACT: The Hippo pathway is crucial in organ size control, and its dysregulation contributes to tumorigenesis. However, upstream signals that regulate the mammalian Hippo pathway have remained elusive. Here, we report that the Hippo pathway is regulated by G-protein-coupled receptor (GPCR) signaling. Serum-borne lysophosphatidic acid (LPA) and sphingosine 1-phosphophate (S1P) act through G12/13-coupled receptors to inhibit the Hippo pathway kinases Lats1/2, thereby activating YAP and TAZ transcription coactivators, which are oncoproteins repressed by Lats1/2. YAP and TAZ are involved in LPA-induced gene expression, cell migration, and proliferation. In contrast, stimulation of Gs-coupled receptors by glucagon or epinephrine activates Lats1/2 kinase activity, thereby inhibiting YAP function. Thus, GPCR signaling can either activate or inhibit the Hippo-YAP pathway depending on the coupled G protein. Our study identifies extracellular diffusible signals that modulate the Hippo pathway and also establishes the Hippo-YAP pathway as a critical signaling branch downstream of GPCR.
    Cell 08/2012; 150(4):780-91. DOI:10.1016/j.cell.2012.06.037 · 33.12 Impact Factor
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