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MOB1 Mediated Phospho-recognition in the Core Mammalian Hippo Pathway

The Hippo tumor suppressor pathway regulates organ size and tissue homoeostasis in response to diverse signaling inputs. The core of the pathway consists of a short kinase cascade: MST1 and MST2 phosphorylate and activate LATS1 and LATS2, which in turn phosphorylate and inactivate key transcriptiona...

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Autores principales: Couzens, Amber L., Xiong, Shawn, Knight, James D. R., Mao, Daniel Y., Guettler, Sebastian, Picaud, Sarah, Kurinov, Igor, Filippakopoulos, Panagis, Sicheri, Frank, Gingras, Anne-Claude
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Biochemistry and Molecular Biology 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5461540/
https://www.ncbi.nlm.nih.gov/pubmed/28373298
http://dx.doi.org/10.1074/mcp.M116.065490
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author Couzens, Amber L.
Xiong, Shawn
Knight, James D. R.
Mao, Daniel Y.
Guettler, Sebastian
Picaud, Sarah
Kurinov, Igor
Filippakopoulos, Panagis
Sicheri, Frank
Gingras, Anne-Claude
author_facet Couzens, Amber L.
Xiong, Shawn
Knight, James D. R.
Mao, Daniel Y.
Guettler, Sebastian
Picaud, Sarah
Kurinov, Igor
Filippakopoulos, Panagis
Sicheri, Frank
Gingras, Anne-Claude
author_sort Couzens, Amber L.
collection PubMed
description The Hippo tumor suppressor pathway regulates organ size and tissue homoeostasis in response to diverse signaling inputs. The core of the pathway consists of a short kinase cascade: MST1 and MST2 phosphorylate and activate LATS1 and LATS2, which in turn phosphorylate and inactivate key transcriptional coactivators, YAP1 and TAZ (gene WWTR1). The MOB1 adapter protein regulates both phosphorylation reactions firstly by concurrently binding to the upstream MST and downstream LATS kinases to enable the trans phosphorylation reaction, and secondly by allosterically activating the catalytic function of LATS1 and LATS2 to directly stimulate phosphorylation of YAP and TAZ. Studies of yeast Mob1 and human MOB1 revealed that the ability to recognize phosphopeptide sequences in their interactors, Nud1 and MST2 respectively, was critical to their roles in regulating the Mitotic Exit Network in yeast and the Hippo pathway in metazoans. However, the underlying rules of phosphopeptide recognition by human MOB1, the implications of binding specificity for Hippo pathway signaling, and the generality of phosphopeptide binding function to other human MOB family members remained elusive. Employing proteomics, peptide arrays and biochemical analyses, we systematically examine the phosphopeptide binding specificity of MOB1 and find it to be highly complementary to the substrate phosphorylation specificity of MST1 and MST2. We demonstrate that autophosphorylation of MST1 and MST2 on several threonine residues provides multiple MOB1 binding sites with varying binding affinities which in turn contribute to a redundancy of MST1-MOB1 protein interactions in cells. The crystal structures of MOB1A in complex with two favored phosphopeptide sites in MST1 allow for a full description of the MOB1A phosphopeptide-binding consensus. Lastly, we show that the phosphopeptide binding properties of MOB1A are conserved in all but one of the seven MOB family members in humans, thus providing a starting point for uncovering their elusive cellular functions.
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spelling pubmed-54615402017-06-14 MOB1 Mediated Phospho-recognition in the Core Mammalian Hippo Pathway Couzens, Amber L. Xiong, Shawn Knight, James D. R. Mao, Daniel Y. Guettler, Sebastian Picaud, Sarah Kurinov, Igor Filippakopoulos, Panagis Sicheri, Frank Gingras, Anne-Claude Mol Cell Proteomics Research The Hippo tumor suppressor pathway regulates organ size and tissue homoeostasis in response to diverse signaling inputs. The core of the pathway consists of a short kinase cascade: MST1 and MST2 phosphorylate and activate LATS1 and LATS2, which in turn phosphorylate and inactivate key transcriptional coactivators, YAP1 and TAZ (gene WWTR1). The MOB1 adapter protein regulates both phosphorylation reactions firstly by concurrently binding to the upstream MST and downstream LATS kinases to enable the trans phosphorylation reaction, and secondly by allosterically activating the catalytic function of LATS1 and LATS2 to directly stimulate phosphorylation of YAP and TAZ. Studies of yeast Mob1 and human MOB1 revealed that the ability to recognize phosphopeptide sequences in their interactors, Nud1 and MST2 respectively, was critical to their roles in regulating the Mitotic Exit Network in yeast and the Hippo pathway in metazoans. However, the underlying rules of phosphopeptide recognition by human MOB1, the implications of binding specificity for Hippo pathway signaling, and the generality of phosphopeptide binding function to other human MOB family members remained elusive. Employing proteomics, peptide arrays and biochemical analyses, we systematically examine the phosphopeptide binding specificity of MOB1 and find it to be highly complementary to the substrate phosphorylation specificity of MST1 and MST2. We demonstrate that autophosphorylation of MST1 and MST2 on several threonine residues provides multiple MOB1 binding sites with varying binding affinities which in turn contribute to a redundancy of MST1-MOB1 protein interactions in cells. The crystal structures of MOB1A in complex with two favored phosphopeptide sites in MST1 allow for a full description of the MOB1A phosphopeptide-binding consensus. Lastly, we show that the phosphopeptide binding properties of MOB1A are conserved in all but one of the seven MOB family members in humans, thus providing a starting point for uncovering their elusive cellular functions. The American Society for Biochemistry and Molecular Biology 2017-06 2017-04-03 /pmc/articles/PMC5461540/ /pubmed/28373298 http://dx.doi.org/10.1074/mcp.M116.065490 Text en © 2017 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version free via Creative Commons CC-BY license (http://creativecommons.org/licenses/by/4.0) .
spellingShingle Research
Couzens, Amber L.
Xiong, Shawn
Knight, James D. R.
Mao, Daniel Y.
Guettler, Sebastian
Picaud, Sarah
Kurinov, Igor
Filippakopoulos, Panagis
Sicheri, Frank
Gingras, Anne-Claude
MOB1 Mediated Phospho-recognition in the Core Mammalian Hippo Pathway
title MOB1 Mediated Phospho-recognition in the Core Mammalian Hippo Pathway
title_full MOB1 Mediated Phospho-recognition in the Core Mammalian Hippo Pathway
title_fullStr MOB1 Mediated Phospho-recognition in the Core Mammalian Hippo Pathway
title_full_unstemmed MOB1 Mediated Phospho-recognition in the Core Mammalian Hippo Pathway
title_short MOB1 Mediated Phospho-recognition in the Core Mammalian Hippo Pathway
title_sort mob1 mediated phospho-recognition in the core mammalian hippo pathway
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5461540/
https://www.ncbi.nlm.nih.gov/pubmed/28373298
http://dx.doi.org/10.1074/mcp.M116.065490
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