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 - Publisher Copyright:
© 2024 The Author(s)
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
UR - http://www.scopus.com/inward/record.url?scp=85214684125&partnerID=8YFLogxK
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 - 2211-1247
VL - 44
JO - Cell Reports
JF - Cell Reports
IS - 1
M1 - 115196
ER -