An Inducible Translocation Strategy to Rapidly Activate and Inhibit Small GTPase Signaling Pathways

Nat Methods. 2005 Jun;2(6):415-8. doi: 10.1038/nmeth763.

Abstract

We made substantial advances in the implementation of a rapamycin-triggered heterodimerization strategy. Using molecular engineering of different targeting and enzymatic fusion constructs and a new rapamycin analog, Rho GTPases were directly activated or inactivated on a timescale of seconds, which was followed by pronounced cell morphological changes. As signaling processes often occur within minutes, such rapid perturbations provide a powerful tool to investigate the role, selectivity and timing of Rho GTPase-mediated signaling processes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism*
  • Enzyme Inhibitors
  • Kinetics
  • Mice
  • NIH 3T3 Cells
  • Protein Engineering / methods*
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction / drug effects
  • Signal Transduction / physiology*
  • Sirolimus / pharmacology*
  • Translocation, Genetic / genetics
  • rho GTP-Binding Proteins / antagonists & inhibitors*
  • rho GTP-Binding Proteins / metabolism*

Substances

  • Enzyme Inhibitors
  • Recombinant Fusion Proteins
  • rho GTP-Binding Proteins
  • Sirolimus