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Determination of the interactome of Non-Structural Protein12 from Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus with Host Cellular Proteins UsingHigh Throughput Proteomics and Identification of Hsp70 as a Critical Cellular Factor for Virus Biology

ABSTRACT: Porcine reproductive and respiratory syndrome virus (PRRSV) has caused tremendous economic losses and continues to be a serious problem to the swine industry worldwide. Although extensive research has been focused on PRRSV, the structure and function of some viral proteins like nonstructural protein12 (NSP12), which may play important roles in viral replication and production, still remain unknown. In order to better understand the function of NSP12, we investigated the interaction of NSP12 with cellular proteins using quantitative proteomics coupled with an immune-precipitation strategy based on the over expression of an NSP12-EGFP fusion protein in 293T cells. Data analysis identified 112 cellular proteins interacted with NSP12-EGFP with high probability. The majority of those proteinsare nucleic acid binding proteins or chaperones, which are involved in RNA post-transcriptional modification, protein synthesis and cellular assembly and organization.Among them, cellular chaperon Hsp70 was verified to interact with PRRSV NSP12 protein, and inhibition of HSP70 significantly reduced the viral mRNA synthesis and virus replication. Our data suggested that NSP12 could recruit cellular proteins such as HSP70 to maintain its own stability and benefit for the virus replication.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Early Embryonic Cell

DISEASE(S): Porcine Reproductive And Respiratory Syndrome

SUBMITTER: Stuart Armstrong

LAB HEAD: Julian Hiscox

PROVIDER: PXD004003 | Pride | 2016-06-27

REPOSITORIES: Pride

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Determination of the interactome of non-structural protein12 from highly pathogenic porcine reproductive and respiratory syndrome virus with host cellular proteins using high throughput proteomics and identification of HSP70 as a cellular factor for virus replication.

Dong Shan S Liu Long L Wu Weining W Armstrong Stuart D SD Xia Dong D Nan Hao H Hiscox Julian A JA Chen Hongying H

Journal of proteomics 20160617

<h4>Unlabelled</h4>Porcine reproductive and respiratory syndrome virus (PRRSV) has caused tremendous economic losses and continues to be a serious problem to the swine industry worldwide. Although extensive research has been focused on PRRSV, the structure and function of some viral proteins like nonstructural protein12 (NSP12), which may play important roles in viral replication and production, still remain unknown. In order to better understand the function of NSP12, we investigated the intera ...[more]

PMID: 27327135

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			Action	DRS
	F016646.dat	Other
	SA_230514_APMS_C12_1.raw	Raw
	SA_230514_APMS_C12_2.raw	Raw
	SA_230514_APMS_CONTROL_1.raw	Raw
	SA_230514_APMS_CONTROL_2.raw	Raw