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The organization of leukotriene biosynthesis on the nuclear envelope revealed by single molecule localization microscopy and computational analyses

Author

Listed:
  • Angela B Schmider
  • Melissa Vaught
  • Nicholas C Bauer
  • Hunter L Elliott
  • Matthew D Godin
  • Giorgianna E Ellis
  • Peter A Nigrovic
  • Roy J Soberman

Abstract

The initial steps in the synthesis of leukotrienes are the translocation of 5-lipoxygenase (5-LO) to the nuclear envelope and its subsequent association with its scaffold protein 5-lipoxygenase-activating protein (FLAP). A major gap in our understanding of this process is the knowledge of how the organization of 5-LO and FLAP on the nuclear envelope regulates leukotriene synthesis. We combined single molecule localization microscopy with Clus-DoC cluster analysis, and also a novel unbiased cluster analysis to analyze changes in the relationships between 5-LO and FLAP in response to activation of RBL-2H3 cells to generate leukotriene C4. We identified the time-dependent reorganization of both 5-LO and FLAP into higher-order assemblies or clusters in response to cell activation via the IgE receptor. Clus-DoC analysis identified a subset of these clusters with a high degree of interaction between 5-LO and FLAP that specifically correlates with the time course of LTC4 synthesis, strongly suggesting their role in the initiation of leukotriene biosynthesis.

Suggested Citation

  • Angela B Schmider & Melissa Vaught & Nicholas C Bauer & Hunter L Elliott & Matthew D Godin & Giorgianna E Ellis & Peter A Nigrovic & Roy J Soberman, 2019. "The organization of leukotriene biosynthesis on the nuclear envelope revealed by single molecule localization microscopy and computational analyses," PLOS ONE, Public Library of Science, vol. 14(2), pages 1-19, February.
    • Handle: RePEc:plo:pone00:0211943
    DOI: 10.1371/journal.pone.0211943
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    1. Sarah L Veatch & Benjamin B Machta & Sarah A Shelby & Ethan N Chiang & David A Holowka & Barbara A Baird, 2012. "Correlation Functions Quantify Super-Resolution Images and Estimate Apparent Clustering Due to Over-Counting," PLOS ONE, Public Library of Science, vol. 7(2), pages 1-13, February.
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