Interaction network of African swine fever virus structural protein p30 with host proteins

Xiongnan Chen; Xiaojun Chen; Yifan Liang; Sijia Xu; Zhijun Weng; Qi Gao; Zhao Huang; Guihong Zhang; Lang Gong

doi:10.3389/fmicb.2022.971888

Interaction network of African swine fever virus structural protein p30 with host proteins

Front Microbiol. 2022 Aug 15:13:971888. doi: 10.3389/fmicb.2022.971888. eCollection 2022.

Authors

Xiongnan Chen^{1

2

3}, Xiaojun Chen^{1

2

3}, Yifan Liang^{1

2

3}, Sijia Xu^{1

2

3}, Zhijun Weng^{1

3}, Qi Gao^{1

2

3}, Zhao Huang^{1

2

3}, Guihong Zhang^{1

2

3

4

5}, Lang Gong^{1

3

6}

Affiliations

¹ Guangdong Provincial Key Laboratory of Zoonosis Prevention and Control, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
² African Swine Fever Regional Laboratory of China, Guangzhou, China.
³ Research Center for African Swine Fever Prevention and Control, South China Agricultural University, Guangzhou, China.
⁴ National Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou, China.
⁵ Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou, Guangdong, China.
⁶ Key Laboratory of Animal Vaccine Development, Ministry of Agriculture and Rural Affairs, Guangzhou, Guangdong, China.

Abstract

African swine fever virus (ASFV) is a complex nucleocytoplasmic large DNA virus (NCLDV) that causes a lethal hemorrhagic disease that is currently threatening the global pig industry. ASFV structural protein p30 is a membrane phosphoprotein that suggests it may play a regulatory role, possibly in signal transduction. Despite its significance in internalization into host cells, the interaction between p30 and host proteins is relatively unknown. In this study, we describe the application of a DUALmembrane yeast two-hybrid assay to screen a primary porcine alveolar macrophages cDNA library and analyze the interactome of p30 protein. Our data identify seven host cellular proteins (DAB2, RPSA, OAS1, PARP9, CAPG, ARPC5, and VBP1) that putatively interact with the p30. We further verified the interaction between p30 and host proteins by laser confocal microscopy, co-immunoprecipitation, and GST-pulldown assay. To further understand the relationship between host proteins and p30, we drew the interaction network diagram and analyzed the functional enrichment of each host protein. Enrichment analysis of Gene Ontology and Kyoto Encyclopedia of Genes and Genomes indicated that host proteins were mainly related to endocytosis, actin cytoskeleton regulation, and innate immunity. Collectively, we identified the interaction between p30 and host cell protein using a membrane protein yeast two-hybrid system, which increases our knowledge of the interaction between ASFV and the host and informs future research on antiviral strategies.

Keywords: African swine fever virus; endocytosis; innate immunity; membrane protein interaction; p30.