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

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, Laura Martin-Sancho, Sumit Chanda, Xinming Li, Steve Bensenger, Nevan J. Krogan and Ren Sun ()
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Yushen Du: University of California
Judd F. Hultquist: University of California, San Francisco
Quan Zhou: University of California
Anders Olson: University of California
Yenwen Tseng: University of California
Tian-hao Zhang: 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
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
Nevan J. Krogan: University of California, San Francisco
Ren Sun: University of California

Nature Communications, 2020, vol. 11, issue 1, 1-13

Abstract: 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.

Date: 2020
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DOI: 10.1038/s41467-020-16140-9

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