TY - JOUR
T1 - Probing the functional constraints of influenza A virus NEP by deep mutational scanning
AU - Teo, Qi Wen
AU - Wang, Yiquan
AU - Lv, Huibin
AU - Oade, Michael S.
AU - Mao, Kevin J.
AU - Tan, Timothy J.C.
AU - Huan, Yang Wei
AU - Rivera-Cardona, Joel
AU - Shao, Evan K.
AU - Choi, Danbi
AU - Wang, Chaoyang
AU - Dargani, Zahra Tavakoli
AU - Brooke, Christopher B.
AU - te Velthuis, Aartjan J.W.
AU - Wu, Nicholas C.
N1 - We thank the Roy J. Carver Biotechnology Center at the University of Illinois Urbana-Champaign for assistance with next-generation sequencing and RNA sequencing. We are grateful to the Institute of Genomic Biology Core Facility at the University of Illinois Urbana-Champaign for providing access to the LSM880 microscope system. This work was supported by the Searle Scholars Program (N.C.W.) and National Institutes of Health (NIH) grant nos. R01 AI165475 (N.C.W.), DP2 AI175474-01 (A.J.W.t.V.), and R01 AI139246 (C.B.B.).
PY - 2025/1/28
Y1 - 2025/1/28
N2 - The influenza A virus nuclear export protein (NEP) is a multifunctional protein that is essential for the viral life cycle and has very high sequence conservation. However, since the open reading frame of NEP largely overlaps with that of another influenza viral protein, non-structural protein 1, it is difficult to infer the functional constraints of NEP based on sequence conservation analysis. In addition, the N-terminal of NEP is structurally disordered, which further complicates the understanding of its function. Here, we systematically measure the replication fitness effects of >1,800 mutations of NEP. Our results show that the N-terminal domain has high mutational tolerance. Additional experiments show that N-terminal domain mutations affect viral transcription and replication dynamics, host cellular responses, and mammalian adaptation of avian influenza virus. Overall, our study not only advances the functional understanding of NEP but also provides insights into its evolutionary constraints.
AB - The influenza A virus nuclear export protein (NEP) is a multifunctional protein that is essential for the viral life cycle and has very high sequence conservation. However, since the open reading frame of NEP largely overlaps with that of another influenza viral protein, non-structural protein 1, it is difficult to infer the functional constraints of NEP based on sequence conservation analysis. In addition, the N-terminal of NEP is structurally disordered, which further complicates the understanding of its function. Here, we systematically measure the replication fitness effects of >1,800 mutations of NEP. Our results show that the N-terminal domain has high mutational tolerance. Additional experiments show that N-terminal domain mutations affect viral transcription and replication dynamics, host cellular responses, and mammalian adaptation of avian influenza virus. Overall, our study not only advances the functional understanding of NEP but also provides insights into its evolutionary constraints.
KW - CP: Microbiology
KW - CP: Molecular biology
KW - NEP
KW - NS1
KW - cellular apoptosis
KW - evolutionary constraints
KW - influenza virus
KW - innate immune response
KW - mammalian adaptation of avian influenza virus
KW - nuclear export protein
KW - the transcription-to-replication switch
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UR - http://www.scopus.com/inward/citedby.url?scp=85214684125&partnerID=8YFLogxK
U2 - 10.1016/j.celrep.2024.115196
DO - 10.1016/j.celrep.2024.115196
M3 - Article
C2 - 39817904
AN - SCOPUS:85214684125
SN - 2639-1856
VL - 44
JO - Cell Reports
JF - Cell Reports
IS - 1
M1 - 115196
ER -