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LPA-induced mutually exclusive subcellular localization of active RhoA and Arp2 mRNA revealed by sequential FRET and

Lisa A Mingle1, Ghislain Bonamy, Margarida Barroso

  • 1Center for Cell Biology and Cancer Research, Albany Medical College, 47 New Scotland Avenue, Albany, NY 12208, USA.

Histochemistry and Cell Biology
|April 15, 2009
PubMed

Insights

Lysophosphatidic acid (LPA) induces Arp2 mRNA localization to cell protrusions via the RhoA-ROCK pathway. This study reveals the crucial role of localized RhoA activation in regulating Arp2/3 complex mRNA transport.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • The Arp2/3 complex is essential for actin nucleation and cell motility.
  • Previous work showed Arp2/3 complex subunits' mRNA localizes to fibroblast protrusions.
  • The signaling pathway governing this mRNA localization was previously unknown.

Purpose of the Study:

  • To identify the signaling pathway regulating Arp2/3 complex mRNA localization.
  • To investigate the role of localized RhoA activation in Arp2 mRNA localization.
  • To develop a method for simultaneously detecting protein activity and mRNA localization.

Main Methods:

  • Fluorescence resonance energy transfer (FRET) to visualize active RhoA.
  • Fluorescence in situ hybridization (FISH) to detect Arp2 mRNA.
  • Sequential FRET and FISH in the same fibroblast cells stimulated with lysophosphatidic acid (LPA).

Main Results:

  • Lysophosphatidic acid (LPA) was identified as a potent inducer of Arp2 mRNA localization to protrusions.
  • The RhoA-ROCK pathway mediates LPA-induced Arp2 mRNA localization.
  • LPA stimulation resulted in mutually exclusive localization of active RhoA and Arp2 mRNA in approximately twice as many cells compared to serum-free conditions.
  • Demonstrated the importance of spatially restricted RhoA activation for Arp2 mRNA localization.

Conclusions:

  • Localized RhoA activation is critical for Arp2/3 complex mRNA localization to cell protrusions.
  • This study provides novel insights into RhoA's regulation of Arp2/3 complex mRNA localization.
  • The combined FRET-FISH technique offers a powerful new method for studying localized protein activity and nucleic acid distribution in single cells.

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