14-3-3 proteins interact with a hybrid prenyl-phosphorylation motif to inhibit G proteins

Philippe Riou, Svend Kjær, Ritu Garg, Andrew Purkiss, Roger George, Robert J Cain, Ganka Bineva, Nicolas Reymond, Brad McColl, Andrew J Thompson, Nicola O'Reilly, Neil Q McDonald, Peter J Parker, Anne J Ridley

Research output: Contribution to journalArticlepeer-review

Abstract

Signaling through G proteins normally involves conformational switching between GTP- and GDP-bound states. Several Rho GTPases are also regulated by RhoGDI binding and sequestering in the cytosol. Rnd proteins are atypical constitutively GTP-bound Rho proteins, whose regulation remains elusive. Here, we report a high-affinity 14-3-3-binding site at the C terminus of Rnd3 consisting of both the Cys241-farnesyl moiety and a Rho-associated coiled coil containing protein kinase (ROCK)-dependent Ser240 phosphorylation site. 14-3-3 binding to Rnd3 also involves phosphorylation of Ser218 by ROCK and/or Ser210 by protein kinase C (PKC). The crystal structure of a phosphorylated, farnesylated Rnd3 peptide with 14-3-3 reveals a hydrophobic groove in 14-3-3 proteins accommodating the farnesyl moiety. Functionally, 14-3-3 inhibits Rnd3-induced cell rounding by translocating it from the plasma membrane to the cytosol. Rnd1, Rnd2, and geranylgeranylated Rap1A interact similarly with 14-3-3. In contrast to the canonical GTP/GDP switch that regulates most Ras superfamily members, our results reveal an unprecedented mechanism for G protein inhibition by 14-3-3 proteins.
Original languageEnglish
Pages (from-to)640-53
Number of pages14
JournalCell
Volume153
Issue number3
DOIs
Publication statusPublished - 25 Apr 2013

Keywords

  • 14-3-3 Proteins
  • Amino Acid Sequence
  • Animals
  • COS Cells
  • Cell Membrane
  • Cercopithecus aethiops
  • Crystallography, X-Ray
  • Cytosol
  • Humans
  • Models, Molecular
  • Molecular Sequence Data
  • Phosphorylation
  • Prenylation
  • Protein Interaction Domains and Motifs
  • rho GTP-Binding Proteins

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