Phosphorylation regulates the nucleocytoplasmic distribution of kinase suppressor of Ras

Jennifer A. Brennan, Deanna J. Volle, Oleg V. Chaika, Robert E Lewis

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

KSR (kinase suppressor of Ras) has been proposed as a molecular scaffold regulating the Raf/MEK/ERK kinase cascade. KSR is phosphorylated on multiple phosphorylation sites by associated kinases. To identify potential mechanisms used by KSR to regulate ERK activation, green fluorescent protein was fused to intact and mutated KSR constructs lacking specific phosphorylation sites, and the subcellular distribution of each construct was observed in live cells. Mutation of a subset of KSR phosphorylation sites caused the redistribution of KSR to the nucleus. To determine whether intact KSR is normally imported to the nucleus, REF-52 fibroblasts expressing KSR were treated with 10 nM leptomycin B, which inhibits Crml-dependent nuclear export. KSR accumulated in the nucleus within 2 h of treatment with leptomycin B, suggesting that KSR cycles continuously through the nucleus. Nuclear import of KSR was blocked by mutations that inhibit the interaction of KSR with MEK. Coexpression of fluorescent forms of KSR and MEK in cells revealed that each protein promoted the localization of the other in the cytoplasm. These data indicate that the subcellular distribution of KSR is dynamically regulated through phosphorylation and MEK interaction in a manner that may affect signaling through ERK.

Original languageEnglish (US)
Pages (from-to)5369-5377
Number of pages9
JournalJournal of Biological Chemistry
Volume277
Issue number7
DOIs
StatePublished - Feb 15 2002

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Phosphorylation
Mitogen-Activated Protein Kinase Kinases
Cell Nucleus Active Transport
KSR-1 protein kinase
Phosphotransferases
MAP Kinase Kinase Kinases
Mutation
Fibroblasts
Green Fluorescent Proteins
Scaffolds

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Phosphorylation regulates the nucleocytoplasmic distribution of kinase suppressor of Ras. / Brennan, Jennifer A.; Volle, Deanna J.; Chaika, Oleg V.; Lewis, Robert E.

In: Journal of Biological Chemistry, Vol. 277, No. 7, 15.02.2002, p. 5369-5377.

Research output: Contribution to journalArticle

Brennan, Jennifer A. ; Volle, Deanna J. ; Chaika, Oleg V. ; Lewis, Robert E. / Phosphorylation regulates the nucleocytoplasmic distribution of kinase suppressor of Ras. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 7. pp. 5369-5377.
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