TY - JOUR
T1 - mRNA display with library of even-distribution reveals cellular interactors of influenza virus NS1
AU - Du, Yushen
AU - Hultquist, Judd F.
AU - Zhou, Quan
AU - Olson, Anders
AU - Tseng, Yenwen
AU - Zhang, Tian hao
AU - Hong, Mengying
AU - Tang, Kejun
AU - Chen, Liubo
AU - Meng, Xiangzhi
AU - McGregor, Michael J.
AU - Dai, Lei
AU - Gong, Danyang
AU - Martin-Sancho, Laura
AU - Chanda, Sumit
AU - Li, Xinming
AU - Bensenger, Steve
AU - Krogan, Nevan J.
AU - Sun, Ren
N1 - Funding Information:
Funding support of NIH PO1 CA177322 to R.S., S.C., and N.K. and NIH U19AI135972.
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - 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.
AB - 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.
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U2 - 10.1038/s41467-020-16140-9
DO - 10.1038/s41467-020-16140-9
M3 - Article
C2 - 32415096
AN - SCOPUS:85084787651
VL - 11
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
IS - 1
M1 - 2449
ER -