Abstract: Wang et al. BMC Microbiology
(2025) 25:474
https://doi.org/10.1186/s12866-025-04226-7
BMC Microbiology
Open Access
RESEARCH
Screening and identification of host factors
interacting with the nucleocapsid protein
of SARS-CoV-2 omicron variant using the yeast
two-hybrid system
Chunhua Wang1,2†, Tingyu Yu1†, Moyue Xie1,3†, Dongang Zhan1,3†, Shuaijie Zhang1†, Wenqi Chai1, Jianzhong Zhao2,
Lijuan Yin4, Yang Yang5*, Liang Shen1* and Ying Xia1*
Abstract
Background The nucleocapsid protein (N protein) of SARS-CoV-2 is highly conserved in viral evolution and serves
as the primary structural protein in viral infection, being the most abundant in viral particles. The N protein is highly
immunogenic and plays a key role in the processes of viral infection and replication, making it of significant research
value in both basic studies and clinical applications.
Results To further investigate the functions of SARS-CoV-2 N protein, the Matchmaker Gold Yeast Two-Hybrid System
was used to identify potential interacting partners of the N protein in human peripheral blood mononuclear cells
(PBMCs). Through this approach, we identified 11 host proteins that might interact with the SARS-CoV-2 N protein.
We further validated the interaction between the N protein and two host proteins, RNF2 and ARL15, which showed
the highest positive clone rates at the cellular level. We also predicted the critical amino acid residues mediating the
interaction of the N protein with RNF2 or ARL15. Additionally, we explored the impact of these two host proteins
on coronavirus replication. Functional analysis of all the 11 host proteins revealed their involvement in ribosome
biogenesis, antigen processing and presentation, as well as various signaling pathways such as JAK-STAT.
Conclusions This study further enriches the understanding of interactions between the SARS-CoV-2 N protein
and host, providing important theoretical insights for the deeper understanding of viral pathogenesis and the
development of antiviral strategies.
Keywords SARS-CoV-2, Nucleocapsid, RNF2, ARL15, Yeast two-hybrid system
†
Chunhua Wang, Tingyu Yu, Moyue Xie, Dongang Zhan and Shuaijie
Zhang have contributed equally to this work.
*Correspondence:
Yang Yang
young@mail.sustech.edu.cn
Liang Shen
shenliang.0829@163.com
Ying Xia
xiaying0437@163.com
Full list of author information is available at the end of the article
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