Ras-Sensitive IMP Modulation of the Raf/MEK/ERK Cascade Through KSR1

Sharon A. Matheny, Michael A. White

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The E3 ubiquitin ligase IMP (impedes mitogenic signal propagation) was isolated as a novel Ras effector that negatively regulates ERK1/2 activation. Current evidence suggests that IMP limits the functional assembly of Raf/MEK complexes by inactivation of the KSR1 adaptor/scaffold protein. Interaction with Ras-GTP stimulates IMP autoubiquitination to relieve limitations on KSR function. The elevated sensitivity of IMP-depleted cells to ERK1/2 pathway activation suggests IMP acts as a signal threshold regulator by imposing reversible restrictions on the assembly of functional Raf/MEK/ERK kinase modules. These observations challenge commonly held concepts of signal transmission by Ras to the MAPK pathway and provide evidence for the role of amplitude modulation in tuning cellular responses to ERK1/2 pathway engagement. Here we describe details of the methods, including RNA interference, ubiquitin ligase assays, and protein complex analysis, that can be used to display the Ras-sensitive contribution of IMP to KSR-dependent modulation of the Raf/MEK/ERK pathway.

Original languageEnglish (US)
Pages (from-to)237-247
Number of pages11
JournalMethods in Enzymology
Volume407
DOIs
StatePublished - 2005

Fingerprint

MAP Kinase Signaling System
Mitogen-Activated Protein Kinase Kinases
Modulation
Ubiquitin-Protein Ligase Complexes
Chemical activation
MAP Kinase Kinase Kinases
Ubiquitin-Protein Ligases
Amplitude modulation
RNA Interference
Guanosine Triphosphate
Scaffolds
Assays
Phosphotransferases
Tuning
RNA
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Ras-Sensitive IMP Modulation of the Raf/MEK/ERK Cascade Through KSR1. / Matheny, Sharon A.; White, Michael A.

In: Methods in Enzymology, Vol. 407, 2005, p. 237-247.

Research output: Contribution to journalArticle

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