Identification of host cellular proteins that interact with the M protein of a highly pathogenic porcine reproductive and respiratory syndrome virus vaccine strain

Qian Wang; Yanwei Li; Hong Dong; Li Wang; Jinmei Peng; Tongqing An; Xufu Yang; Zhijun Tian; Xuehui Cai

doi:10.1186/s12985-017-0700-1

Identification of host cellular proteins that interact with the M protein of a highly pathogenic porcine reproductive and respiratory syndrome virus vaccine strain

Virol J. 2017 Feb 22;14(1):39. doi: 10.1186/s12985-017-0700-1.

Authors

Qian Wang¹, Yanwei Li², Hong Dong³, Li Wang³, Jinmei Peng¹, Tongqing An¹, Xufu Yang⁴, Zhijun Tian⁵, Xuehui Cai⁶

Affiliations

¹ Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No.678, Haping street, Xiangfang District, Harbin, 150069, China.
² National Engineering Research Center of Veterinary Biologics, Harbin, 150001, China.
³ Key Laboratory of Animal Epidemiology and Zoonosis of Ministry of Agriculture, College of Veterinary Medicine and State Key Laboratory of Agribiotechnology, China Agricultural University, Beijing, 100193, China.
⁴ North Guangdong Collaborative Innovation and Development Center of Pig Farming and Disease Control, Shaoguan University, Shaoguan, 512005, China.
⁵ Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No.678, Haping street, Xiangfang District, Harbin, 150069, China. tzj@hvri.ac.cn.
⁶ Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No.678, Haping street, Xiangfang District, Harbin, 150069, China. aci139@sina.com.

Abstract

Background: The highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) continues to pose one of the greatest threats to the swine industry. M protein is the most conserved and important structural protein of PRRSV. However, information about the host cellular proteins that interact with M protein remains limited.

Methods: Host cellular proteins that interact with the M protein of HP-PRRSV were immunoprecipitated from MARC-145 cells infected with PRRSV HuN4-F112 using the M monoclonal antibody (mAb). The differentially expressed proteins were identified by LC-MS/MS. The screened proteins were used for bioinformatics analysis including Gene Ontology, the interaction network, and the enriched KEGG pathways. Some interested cellular proteins were validated to interact with M protein by CO-IP.

Results: The PRRSV HuN4-F112 infection group had 10 bands compared with the control group. The bands included 219 non-redundant cellular proteins that interact with M protein, which were identified by LC-MS/MS with high confidence. The gene ontology and Kyoto encyclopedia of genes and genomes (KEGG) pathway bioinformatic analyses indicated that the identified proteins could be assigned to several different subcellular locations and functional classes. Functional analysis of the interactome profile highlighted cellular pathways associated with protein translation, infectious disease, and signal transduction. Two interested cellular proteins-nuclear factor of activated T cells 45 kDa (NF45) and proliferating cell nuclear antigen (PCNA)-that could interact with M protein were validated by Co-IP and confocal analyses.

Conclusions: The interactome data between PRRSV M protein and cellular proteins were identified and contribute to the understanding of the roles of M protein in the replication and pathogenesis of PRRSV. The interactome of M protein will aid studies of virus/host interactions and provide means to decrease the threat of PRRSV to the swine industry in the future.

Keywords: Bioinformatics; HP-PRRSV; Host cellular proteins; Interaction; M protein.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line
Chromatography, Liquid
Haplorhini
Host-Pathogen Interactions*
Immunoprecipitation
Porcine respiratory and reproductive syndrome virus / physiology*
Protein Interaction Mapping*
Swine
Tandem Mass Spectrometry
Viral Matrix Proteins / metabolism*

Substances

M protein, porcine reproductive and respiratory syndrome virus
Viral Matrix Proteins