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Phospholipase Cγ/diacylglycerol‐dependent activation of β2‐chimaerin restricts EGF‐induced Rac signaling

HongBin Wang, Chengfeng Yang, Federico Coluccio Leskow, Jing Sun, Bertram Canagarajah, James H Hurley, Marcelo G Kazanietz

Author Affiliations

  1. HongBin Wang1,
  2. Chengfeng Yang1,
  3. Federico Coluccio Leskow1,
  4. Jing Sun1,
  5. Bertram Canagarajah2,
  6. James H Hurley2 and
  7. Marcelo G Kazanietz*,1
  1. 1 Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, PA, USA
  2. 2 Laboratory of Molecular Biology, NIDDK, NIH, Bethesda, MD, USA
  1. *Corresponding author. Department of Pharmacology, University of Pennsylvania School of Medicine, 816 BRB II/III, 421 Curie Blvd., Philadelphia, PA 19104‐6160, USA. Tel.: +1 215 898 0253; Fax: +1 215 573 9004; E-mail: marcelo{at}spirit.gcrc.upenn.edu
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Abstract

Although receptor‐mediated regulation of small G‐proteins and the cytoskeleton is intensively studied, the mechanisms for attenuation of these signals are poorly understood. In this study, we have identified the Rac‐GAP β2‐chimaerin as an effector of the epidermal growth factor receptor (EGFR) via coupling to phospholipase Cγ (PLCγ) and generation of the lipid second messenger diacylglycerol (DAG). EGF redistributes β2‐chimaerin to promote its association with the small GTPase Rac1 at the plasma membrane, as determined by FRET. This relocalization and association with Rac1 were impaired by disruption of the β2‐chimaerin C1 domain as well as by PLCγ1 RNAi, thus defining β2‐chimaerin as a novel DAG effector. On the other hand, GAP‐deficient β2‐chimaerin mutants show enhanced translocation and sustained Rac1 association in the FRET assays. Remarkably, RNAi depletion of β2‐chimaerin significantly extended the duration of Rac activation by EGF, suggesting that β2‐chimaerin serves as a mechanism that self‐limits Rac activity in response to EGFR activation. Our results represent the first direct evidence of divergence in DAG signaling downstream of a tyrosine‐kinase receptor via a PKC‐independent mechanism.

  • Received October 17, 2005.
  • Accepted March 27, 2006.
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