High-Content Screening Microscopy Identifies Novel Proteins With a Putative Role in Secretory Membrane Traffic

  1. Vytaute Starkuviene1,3,
  2. Urban Liebel1,3,
  3. Jeremy C. Simpson1,
  4. Holger Erfle1,
  5. Annemarie Poustka2,
  6. Stefan Wiemann2, and
  7. Rainer Pepperkok1,4
  1. 1Cell Biology and Biophysics Programme, European Molecular Biology Laboratory (EMBL), 69117 Heidelberg, Germany
  2. 2Molecular Genome Analysis, German Cancer Research Centre, 69120 Heidelberg, Germany

Abstract

Here we describe the establishment of microscope-based functional screening assays in intact cells that allow us to systematically identify new proteins involved in secretory membrane traffic, and proteins that can influence the integrity of the Golgi complex. We were able to identify 20 new proteins that affected either secretory transport, Golgi morphology, or both, when overexpressed in cells. Control experiments with human orthologs to yeast proteins with a role in membrane traffic, or already well characterized mammalian regulators of the secretory pathway, confirmed the specificity and significance of our results. Proteins localized to the Golgi complex or endoplasmic reticulum (ER) showed preferential interference in our assays. Bioinformatic analysis of the new proteins interfering with membrane traffic and/or Golgi integrity revealed broad functional variety, but demonstrated a bias towards proteins with predicted coiled-coil domains and repeat structures. Extending our approach to a much larger set of novel proteins in the future will be an important step toward a more comprehensive understanding of the molecular basis of the secretory pathway. It will also serve as an example for similar microscope-based screens addressing different biological questions.

Footnotes

  • [Supplemental material is available online at www.genome.org.]

  • Article and publication are at http://www.genome.org/cgi/doi/10.1101/gr.2658304.

  • 3 These authors contributed equally to this work.

  • 4 Corresponding author. E-MAIL pepperkok{at}embl.de; FAX 0049 6221 387 306.

    • Accepted July 29, 2004.
    • Received April 8, 2004.
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