Araf kinase antagonizes Nodal-Smad2 activity in mesendoderm development by directly phosphorylating the Smad2 linker region

Smad2/3-mediated transforming growth factor β signalling and the Ras-Raf-Mek-Erk cascade have important roles in stem cell and development and tissue homeostasis. However, it remains unknown whether Raf kinases directly crosstalk with Smad2/3 signalling and how this would regulate embryonic development. Here we show that Araf antagonizes mesendoderm induction and patterning activity of Nodal/Smad2 signals in vertebrate embryos by directly inhibiting Smad2 signalling. Knockdown of araf in zebrafish embryos leads to an increase of activated Smad2 with a decrease in linker phosphorylation; consequently, the embryos have excess mesendoderm precursors and are dorsalized. Mechanistically, Araf physically binds to and phosphorylates Smad2 in the linker region with S253 being indispensable in a Mek/Erk-independent manner, thereby attenuating Smad2 signalling by accelerating degradation of activated Smad2. Our findings open avenues for investigating the potential significance of Raf regulation of transforming growth factor β signalling in versatile biological and pathological processes in the future. TGF-β signalling through SMAD transcription factors has been implicated in embryonic mesendoderm development. Liu and colleagues reveal that the Ras-regulated kinase Raf prevents excessive mesendoderm formation by phosphorylating and promoting the degradation of SMAD2.

• Publication year

  2013

• Venue

  Nature Communications

• Publication date

  2013-04-16

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1038/ncomms2762PMID 23591895PMCID 3644095
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• PPM1A Functions as a Smad Phosphatase to Terminate TGFβ Signaling.

  2016cited by this paper
• Constitutive Smad linker phosphorylation in melanoma: a mechanism of resistance to transforming growth factor‐β‐mediated growth inhibition

  2011cited by this paper
• Graded Nodal/Activin Signaling Titrates Conversion of Quantitative Phospho-Smad2 Levels into Qualitative Embryonic Stem Cell Fate Decisions

  2011cited by this paper
• Mechanistic principles of RAF kinase signaling

  2011cited by this paper
• Global Identification of SMAD2 Target Genes Reveals a Role for Multiple Co-regulatory Factors in Zebrafish Early Gastrulas

  2011cited by this paper
• TGF-β stimulates biglycan synthesis via p38 and ERK phosphorylation of the linker region of Smad2

  2010cited by this paper
• Genome-wide mapping of SMAD target genes reveals the role of BMP signaling in embryonic stem cell fate determination.

  2010cited by this paper
• Smad2 mediates Activin/Nodal signaling in mesendoderm differentiation of mouse embryonic stem cells

  2010cited by this paper
• Tgf-beta superfamily signaling in embryonic development and homeostasis.

  2009cited by this paper
• Phospho-control of TGF-β superfamily signaling

  2009cited by this paper
• Rock2 controls TGFβ signaling and inhibits mesoderm induction in zebrafish embryos

  2009cited by this paper
• Smad2/3 activities are required for induction and patterning of the neuroectoderm in zebrafish.

  2009cited by this paper
• Distinct functions for ERK1 and ERK2 in cell migration processes during zebrafish gastrulation.

  2008cited by this paper
• smad2 and smad3 Are Required for Mesendoderm Induction by Transforming Growth Factor-β/Nodal Signals in Zebrafish*

  2008cited by this paper
• TGFbeta in Cancer.

  2008cited by this paper
• Integrating patterning signals: Wnt/GSK3 regulates the duration of the BMP/Smad1 signal.

  2007cited by this paper
• Balancing BMP signaling through integrated inputs into the Smad1 linker.

  2007cited by this paper
• Regulation of MAPKs by growth factors and receptor tyrosine kinases.

  2007cited by this paper
• Germline gain-of-function mutations in SOS1 cause Noonan syndrome

  2007cited by this paper
• Germline gain-of-function mutations in RAF1 cause Noonan syndrome

  2007cited by this paper
• TGFβ–SMAD signal transduction: molecular specificity and functional flexibility

  2007cited by this paper
• Nodal signals pattern vertebrate embryos

  2006cited by this paper
• Characterization and expression patterns of the MAPK family in zebrafish.

  2006cited by this paper
• PPM1A functions as a Smad phosphatase to terminate TGFbeta signaling.

  2006cited by this paper
• Default neural induction: neuralization of dissociated Xenopus cells is mediated by Ras/MAPK activation.

  2005cited by this paper
• Dorsal-ventral patterning and neural induction in Xenopus embryos.

  2004cited by this paper
• Prediction of phosphorylation sites using SVMs

  2004cited by this paper
• fgf17b, a novel member of Fgf family, helps patterning zebrafish embryos.

  2004cited by this paper
• The RAF proteins take centre stage

  2004cited by this paper
• Integration of IGF, FGF, and anti-BMP signals via Smad1 phosphorylation in neural induction.

  2003cited by this paper
• Evidence for antagonism of BMP-4 signals by MAP kinase during Xenopus axis determination and neural specification.

  2003cited by this paper
• Endodermal Nodal-related signals and mesoderm induction in Xenopus.

  2000cited by this paper
• Equivalent genetic roles for bmp7/snailhouse and bmp2b/swirl in dorsoventral pattern formation.

  2000cited by this paper
• The zebrafish bonnie and clyde gene encodes a Mix family homeodomain protein that regulates the generation of endodermal precursors.

  2000cited by this paper
• The smad5 mutation somitabun blocks Bmp2b signaling during early dorsoventral patterning of the zebrafish embryo.

  1999cited by this paper
• casanova plays an early and essential role in endoderm formation in zebrafish.

  1999cited by this paper
• The EGF-CFC protein one-eyed pinhead is essential for nodal signaling.

  1999cited by this paper
• A mechanism of repression of TGFbeta/ Smad signaling by oncogenic Ras.

  1999cited by this paper
• Induction of the mesendoderm in the zebrafish germ ring by yolk cell-derived TGF-beta family signals and discrimination of mesoderm and endoderm by FGF.

  1999cited by this paper
• Antivin, a novel and divergent member of the TGFbeta superfamily, negatively regulates mesoderm induction.

  1999cited by this paper
• A Phosphatidylinositol 3-Kinase-dependent Pathway That Differentially Regulates c-Raf and A-Raf*

  1999cited by this paper
• A molecular pathway leading to endoderm formation in zebrafish.

  1999cited by this paper
• Failure of egg cylinder elongation and mesoderm induction in mouse embryos lacking the tumor suppressor smad2.

  1998cited by this paper
• Zebrafish organizer development and germ-layer formation require nodal-related signals

  1998cited by this paper
• Zebrafish nodal-related genes are implicated in axial patterning and establishing left-right asymmetry.

  1998cited by this paper
• Smad2 role in mesoderm formation, left–right patterning and craniofacial development

  1998cited by this paper
• A role for FGF-8 in the dorsoventral patterning of the zebrafish gastrula.

  1997cited by this paper
• The molecular nature of zebrafish swirl: BMP2 function is essential during early dorsoventral patterning.

  1997cited by this paper
• Opposing BMP and EGF signalling pathways converge on the TGF-β family mediator Smad1

  1997cited by this paper
• Identification of a potential regulator of early transcriptional responses to mesoderm inducers in the frog embryo.

  1995cited by this paper
• The expression of a zebrafish gene homologous to Drosophila snail suggests a conserved function in invertebrate and vertebrate gastrulation.

  1993cited by this paper
• Nodal is a novel TGF-β-like gene expressed in the mouse node during gastrulation

  1993cited by this paper
• Nodal is a novel TGF-beta-like gene expressed in the mouse node during gastrulation.

  1993cited by this paper
• Origin and organization of the zebrafish fate map.

  1990cited by this paper

• Phosphoproteomic analysis reveals changes in A-Raf-related protein phosphorylation in response to Toxoplasma gondii infection in porcine macrophages

  2024cites this paper
• Receptor-activated transcription factors and beyond: multiple modes of Smad2/3-dependent transmission of TGF-β signaling.

  2024cites this paper
• A Huluwa phosphorylation switch regulates embryonic axis induction

  2024cites this paper
• Cardiomyocyte intercellular signalling increases oxidative stress and reprograms the global‐ and phospho‐proteome of cardiac fibroblasts

  2023cites this paper
• Smad2Δexon3 and Smad3 have distinct properties in signal transmission leading to TGF-β–induced cell motility

  2022cites this paper
• The phosphorylation of the Smad2/3 linker region by nemo-like kinase regulates TGF-β signaling

  2021cites this paper
• miR-103/107 regulates left-right asymmetry in zebrafish by modulating Kupffer's vesicle development and ciliogenesis.

  2020cites this paper
• Dominant-negative antagonists of the Ras-ERK pathway: DA-Raf and its related proteins generated by alternative splicing of Raf.

  2019cites this paper
• Coordinate regulation of retinoic acid synthesis by pbx genes and fibroblast growth factor signaling by hoxb1b is required for hindbrain patterning and development.

  2018cites this paper
• Molecular regulation of Nodal signaling during mesendoderm formation

  2018cites this paper
• Maternal Ybx1 safeguards zebrafish oocyte maturation and maternal-to-zygotic transition by repressing global translation

  2018influential citation
• Maternal Huluwa dictates the embryonic body axis through β-catenin in vertebrates

  2018cites this paper
• A GEF activity-independent function for nuclear Net1 in Nodal signal transduction and mesendoderm formation

  2017cites this paper
• Alkbh4 and Atrn Act Maternally to Regulate Zebrafish Epiboly

  2017cites this paper
• Identification of SSEA-1 expressing enhanced reprogramming (SEER) cells in porcine embryonic fibroblasts

  2017cites this paper
• Nuclear movement regulated by non-Smad Nodal signaling via JNK is associated with Smad signaling during zebrafish endoderm specification

  2017cites this paper
• Embryonic hematopoiesis in vertebrate somites gives rise to definitive hematopoietic stem cells

  2016cites this paper
• Posttranslational Regulation of Smads.

  2016cites this paper
• Eph/ephrin signaling maintains the boundary of dorsal forerunner cell cluster during morphogenesis of the zebrafish embryonic left-right organizer

  2016cites this paper
• Coordinate involvement of Nodal-dependent inhibition and Wnt-dependent activation in the maintenance of organizer-specific bmp2b in zebrafish.

  2016cites this paper
• Impairment of Cargo Transportation Caused by gbf1 Mutation Disrupts Vascular Integrity and Causes Hemorrhage in Zebrafish Embryos*

  2016cites this paper
• TGFβ1a regulates zebrafish posterior lateral line formation via Smad5 mediated pathway.

  2015cites this paper
• Increased Expression of TGF-β Signaling Components in a Mouse Model of Fibrosis Induced by Submandibular Gland Duct Ligation

  2015cites this paper
• PIAS-like protein Zimp7 is required for the restriction of the zebrafish organizer and mesoderm development.

  2015cites this paper
• A-Raf: A new star of the family of raf kinases

  2015cites this paper
• The Kinase Activity-deficient Isoform of the Protein Araf Antagonizes Ras/Mitogen-activated Protein Kinase (Ras/MAPK) Signaling in the Zebrafish Embryo

  2015cites this paper
• Organizer-derived Bmp2 is required for the formation of a correct Bmp activity gradient during embryonic development

  2014cites this paper
• Maternal Eomesodermin regulates zygotic nodal gene expression for mesendoderm induction in zebrafish embryos.

  2014cites this paper
• Targeting the Wnt pathway in zebrafish as a screening method to identify novel therapeutic compounds

  2014cites this paper