Dissecting the Role of Hsp70 in Beet Black Scorch Virus Infection
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- 英文论文题名：Dissecting the Role of Hsp70 in Beet Black Scorch Virus Infection
- 论文作者：WANG Xiaoling ; CAO Xiu-ling ; LIU Min ; ZHANG Rui-qi ; ZHANG Xin ; WANG Xian-bing ; LI Da-wei ; ZHANG Yong-liang
- 英文论文作者：WANG Xiaoling ; CAO Xiu-ling ; LIU Min ; ZHANG Rui-qi ; ZHANG Xin ; WANG Xian-bing ; LI Da-wei ; ZHANG Yong-liang ; State Key Laboratory of Agro-Biotechnology ; College of Biological Sciences ; China Agricultural University
- 年：2016
- 作者机构：State Key Laboratory of Agro-Biotechnology,College of Biological Sciences,China Agricultural University;
- 英文论文关键词：Beet black scorch virus ; P23 ; CP ; Hsp70 ; Interaction ; Infection
- 会议召开时间：2016-08-05
- 会议录名称：中国植物病理学会2016年学术年会论文集
- 英文会议录名称：Proceedings of the Annual Meeting of Chinese Society for Plant Pathology(2016)
- 语种：英文
- 分类号：S435.663
- 学会代码：ZGVS
- 会议名称：中国植物病理学会2016年学术年会
- 会议地点：中国江苏南京
- 主办单位：中国植物病理学会
- 学会名称：中国植物病理学会
- 页数：1
- 文件大小：52k
- 原文格式：O
- 会议级别：全国
摘要
Many RNA viruses subvert large number of host proteins to complete the viral life cycle,such as genomic RNA replication and viral movement,encapsidation and to interfere with host antiviral responses.In order to find host proteins involved in Beet black scorch virus(BBSV) infection,P23,the auxiliary replication protein,and the coat protein(CP) were fused with a FLAG tag,respectively,followed by agro-infiltration assays.Based upon an affinity purification approach and liquid chromatography-tandem mass spectrometry(LC-MS/MS),we identified heat shock protein70(Hsp70) as a component existing in both complexes that was co-purified with P23 and CP.In this work,we confirmed that Hsp70 interact with both CP and p23 by Co-IP,pull-down,and BiFC,respectively,whereas no interaction was detected between p23 and CP.Furthermore,dual-color trimolecular fluorescence complementation reveals the formation of ternary p23-Hsp70-CP complexes in vivo,which localized to vesicles derived from endoplasmic reticulum.Using qRT-PCR,we revealed that both mRNA and protein level of Hsp70 transcripts increased during BBSV infection.In addition,VIGS or inhibitor down-regulation of Hsp70,leads to the decrement of BBSV genomic RNA and coat protein accumulation.Agrobacterium-mediated expression of p23 leads to hsp70 mRNA up-regulated whereas CP leads to opposite directions.BBSV CP,which acts in RNA encapsidation and long distance movement,also represses viral RNA replication in a dose-dependent manner.Altogether,in order to achieve persistent infection,CP interact with both P23 and Hsp70 to balance replication and encapsidation via redistributed to ER-associated vesicles.Hence,our findings unveiled Hsp70 spatio-temporally regulates the infection of BBSV in N.benthamiana via interacting with different viral components at diverse viral infection stages.
Many RNA viruses subvert large number of host proteins to complete the viral life cycle,such as genomic RNA replication and viral movement,encapsidation and to interfere with host antiviral responses.In order to find host proteins involved in Beet black scorch virus(BBSV) infection,P23,the auxiliary replication protein,and the coat protein(CP) were fused with a FLAG tag,respectively,followed by agro-infiltration assays.Based upon an affinity purification approach and liquid chromatography-tandem mass spectrometry(LC-MS/MS),we identified heat shock protein70(Hsp70) as a component existing in both complexes that was co-purified with P23 and CP.In this work,we confirmed that Hsp70 interact with both CP and p23 by Co-IP,pull-down,and BiFC,respectively,whereas no interaction was detected between p23 and CP.Furthermore,dual-color trimolecular fluorescence complementation reveals the formation of ternary p23-Hsp70-CP complexes in vivo,which localized to vesicles derived from endoplasmic reticulum.Using qRT-PCR,we revealed that both mRNA and protein level of Hsp70 transcripts increased during BBSV infection.In addition,VIGS or inhibitor down-regulation of Hsp70,leads to the decrement of BBSV genomic RNA and coat protein accumulation.Agrobacterium-mediated expression of p23 leads to hsp70 mRNA up-regulated whereas CP leads to opposite directions.BBSV CP,which acts in RNA encapsidation and long distance movement,also represses viral RNA replication in a dose-dependent manner.Altogether,in order to achieve persistent infection,CP interact with both P23 and Hsp70 to balance replication and encapsidation via redistributed to ER-associated vesicles.Hence,our findings unveiled Hsp70 spatio-temporally regulates the infection of BBSV in N.benthamiana via interacting with different viral components at diverse viral infection stages.
Many RNA viruses subvert large number of host proteins to complete the viral life cycle,such as genomic RNA replication and viral movement,encapsidation and to interfere with host antiviral responses.In order to find host proteins involved in Beet black scorch virus(BBSV) infection,P23,the auxiliary replication protein,and the coat protein(CP) were fused with a FLAG tag,respectively,followed by agro-infiltration assays.Based upon an affinity purification approach and liquid chromatography-tandem mass spectrometry(LC-MS/MS),we identified heat shock protein70(Hsp70) as a component existing in both complexes that was co-purified with P23 and CP.In this work,we confirmed that Hsp70 interact with both CP and p23 by Co-IP,pull-down,and BiFC,respectively,whereas no interaction was detected between p23 and CP.Furthermore,dual-color trimolecular fluorescence complementation reveals the formation of ternary p23-Hsp70-CP complexes in vivo,which localized to vesicles derived from endoplasmic reticulum.Using qRT-PCR,we revealed that both mRNA and protein level of Hsp70 transcripts increased during BBSV infection.In addition,VIGS or inhibitor down-regulation of Hsp70,leads to the decrement of BBSV genomic RNA and coat protein accumulation.Agrobacterium-mediated expression of p23 leads to hsp70 mRNA up-regulated whereas CP leads to opposite directions.BBSV CP,which acts in RNA encapsidation and long distance movement,also represses viral RNA replication in a dose-dependent manner.Altogether,in order to achieve persistent infection,CP interact with both P23 and Hsp70 to balance replication and encapsidation via redistributed to ER-associated vesicles.Hence,our findings unveiled Hsp70 spatio-temporally regulates the infection of BBSV in N.benthamiana via interacting with different viral components at diverse viral infection stages.
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