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mRNA display with library of even-distribution reveals cellular interactors of influenza virus NS1

Author

Listed:
  • Yushen Du

    (University of California
    School of Medicine, Zhejiang University)

  • Judd F. Hultquist

    (University of California, San Francisco
    University of California, San Francisco
    J. David Gladstone Institutes
    Northwestern University Feinberg School of Medicine)

  • Quan Zhou

    (University of California)

  • Anders Olson

    (University of California)

  • Yenwen Tseng

    (University of California)

  • Tian-hao Zhang

    (University of California
    University of California)

  • Mengying Hong

    (School of Medicine, Zhejiang University)

  • Kejun Tang

    (School of Medicine, Zhejiang University)

  • Liubo Chen

    (School of Medicine, Zhejiang University)

  • Xiangzhi Meng

    (University of California)

  • Michael J. McGregor

    (University of California, San Francisco
    University of California, San Francisco
    J. David Gladstone Institutes)

  • Lei Dai

    (University of California)

  • Danyang Gong

    (University of California)

  • Laura Martin-Sancho

    (Sanford Burnham Prebys Medical Discovery Institute)

  • Sumit Chanda

    (Sanford Burnham Prebys Medical Discovery Institute)

  • Xinming Li

    (David Geffen School of Medicine at UCLA, L)

  • Steve Bensenger

    (University of California
    University of California)

  • Nevan J. Krogan

    (University of California, San Francisco
    University of California, San Francisco
    J. David Gladstone Institutes)

  • Ren Sun

    (University of California
    University of California)

Abstract

A comprehensive examination of protein-protein interactions (PPIs) is fundamental for the understanding of cellular machineries. However, limitations in current methodologies often prevent the detection of PPIs with low abundance proteins. To overcome this challenge, we develop a mRNA display with library of even-distribution (md-LED) method that facilitates the detection of low abundance binders with high specificity and sensitivity. As a proof-of-principle, we apply md-LED to IAV NS1 protein. Complementary to AP-MS, md-LED enables us to validate previously described PPIs as well as to identify novel NS1 interactors. We show that interacting with FASN allows NS1 to directly regulate the synthesis of cellular fatty acids. We also use md-LED to identify a mutant of NS1, D92Y, results in a loss of interaction with CPSF1. The use of high-throughput sequencing as the readout for md-LED enables sensitive quantification of interactions, ultimately enabling massively parallel experimentation for the investigation of PPIs.

Suggested Citation

  • Yushen Du & Judd F. Hultquist & Quan Zhou & Anders Olson & Yenwen Tseng & Tian-hao Zhang & Mengying Hong & Kejun Tang & Liubo Chen & Xiangzhi Meng & Michael J. McGregor & Lei Dai & Danyang Gong & Laur, 2020. "mRNA display with library of even-distribution reveals cellular interactors of influenza virus NS1," Nature Communications, Nature, vol. 11(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-16140-9
    DOI: 10.1038/s41467-020-16140-9
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