Identification of host cellular proteins LAGE3 and IGFBP6 that interact with orf virus protein ORFV024

Mingjian Long; Yuanyuan Wang; Daxiang Chen; Yong Wang; Ruixue Wang; Daoyuan Gong; Haijian He; Daniel L. Rock; Wenbo Hao; Shuhong Luo

doi:10.1016/j.gene.2018.03.089

Identification of host cellular proteins LAGE3 and IGFBP6 that interact with orf virus protein ORFV024

Mingjian Long, Yuanyuan Wang, Daxiang Chen, Yong Wang, Ruixue Wang, Daoyuan Gong, Haijian He, Daniel L. Rock, Wenbo Hao, Shuhong Luo

Pathobiology

Research output: Contribution to journal › Article › peer-review

Abstract

Objective: Orf virus (ORFV) is the pathogen causing contagious pustular dermatitis in goats, sheep and herdsmen. Evidence has confirmed that ORFV can be used as a preventive and therapeutic immunomodulatory agent in several animal models. Our previous data demonstrated that ORFV024 is able to inhibit activation of the NF-κB signaling pathway and act as an important modulator for early immune responses against viral infection. However, the molecular mechanism by which ORFV024 exerting biological function remains unclear. In the present study, we explored and analyzed the function of host cellular proteins that interact with ORFV024. Methods: The yeast two-hybrid (Y2H) assay was performed to screen proteins interacting with ORFV024 using a cDNA library derived from primary ovine fetal turbinate cells (OFTu). Two of the screened proteins were further confirmed by confocal microscopy, His-tag pull-down assay and CO-Immunoprecipitation (CO-IP) assay. In addition, the ORFV024 interaction network was constructed using the STRING database. Results: In this study, 11 ovine cellular proteins were found to interact with ORFV024. In view of the importance of LAGE3 and IGFBP6 in the ORFV024 functional analysis, we further constructed LAGE3 and IGFBP6 interaction networks. The interactions between ORFV024 and LAGE3 or IGFBP6 were confirmed by confocal microscopy, LAGE3 was further confirmed in the His-tag pull-down assay and CO-IP assay. Conclusions: Our findings indicate that ORFV024 can interact with ovine cellular proteins LAGE3 and IGFBP6.

Original language	English (US)
Pages (from-to)	60-67
Number of pages	8
Journal	Gene
Volume	661
DOIs	https://doi.org/10.1016/j.gene.2018.03.089
State	Published - Jun 30 2018

Keywords

IGFBP6
LAGE3
NF-κB signaling pathway
ORFV024
Orf virus
Yeast two-hybrid system

ASJC Scopus subject areas

Genetics

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10.1016/j.gene.2018.03.089

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@article{5dc5c52a33bb410bbcbf45357628da05,

title = "Identification of host cellular proteins LAGE3 and IGFBP6 that interact with orf virus protein ORFV024",

abstract = "Objective: Orf virus (ORFV) is the pathogen causing contagious pustular dermatitis in goats, sheep and herdsmen. Evidence has confirmed that ORFV can be used as a preventive and therapeutic immunomodulatory agent in several animal models. Our previous data demonstrated that ORFV024 is able to inhibit activation of the NF-κB signaling pathway and act as an important modulator for early immune responses against viral infection. However, the molecular mechanism by which ORFV024 exerting biological function remains unclear. In the present study, we explored and analyzed the function of host cellular proteins that interact with ORFV024. Methods: The yeast two-hybrid (Y2H) assay was performed to screen proteins interacting with ORFV024 using a cDNA library derived from primary ovine fetal turbinate cells (OFTu). Two of the screened proteins were further confirmed by confocal microscopy, His-tag pull-down assay and CO-Immunoprecipitation (CO-IP) assay. In addition, the ORFV024 interaction network was constructed using the STRING database. Results: In this study, 11 ovine cellular proteins were found to interact with ORFV024. In view of the importance of LAGE3 and IGFBP6 in the ORFV024 functional analysis, we further constructed LAGE3 and IGFBP6 interaction networks. The interactions between ORFV024 and LAGE3 or IGFBP6 were confirmed by confocal microscopy, LAGE3 was further confirmed in the His-tag pull-down assay and CO-IP assay. Conclusions: Our findings indicate that ORFV024 can interact with ovine cellular proteins LAGE3 and IGFBP6.",

keywords = "IGFBP6, LAGE3, NF-κB signaling pathway, ORFV024, Orf virus, Yeast two-hybrid system",

author = "Mingjian Long and Yuanyuan Wang and Daxiang Chen and Yong Wang and Ruixue Wang and Daoyuan Gong and Haijian He and Rock, {Daniel L.} and Wenbo Hao and Shuhong Luo",

note = "Funding Information: This study was supported by the National Natural Science Foundation of China [grant numbers: 81773271 and 31672536 ] This project was partially supported by the Foshan University senior talent start fund [grant number: 20161110004 ]; the Foshan University high - level university fund [grant number: 20170131020 ]; and the Agriculture and Food Research Initiative Competitive from the USDA National Institute of Food and Agriculture [grant number: 2012-67015-19289 ]. Funding Information: This study was supported by the National Natural Science Foundation of China [grant numbers: 81773271 and 31672536] This project was partially supported by the Foshan University senior talent start fund [grant number: 20161110004]; the Foshan University high - level university fund [grant number: 20170131020]; and the Agriculture and Food Research Initiative Competitive from the USDA National Institute of Food and Agriculture [grant number: 2012-67015-19289]. Publisher Copyright: {\textcopyright} 2018",

year = "2018",

month = jun,

day = "30",

doi = "10.1016/j.gene.2018.03.089",

language = "English (US)",

volume = "661",

pages = "60--67",

journal = "Gene",

issn = "0378-1119",

publisher = "Elsevier",

}

TY - JOUR

T1 - Identification of host cellular proteins LAGE3 and IGFBP6 that interact with orf virus protein ORFV024

AU - Long, Mingjian

AU - Wang, Yuanyuan

AU - Chen, Daxiang

AU - Wang, Yong

AU - Wang, Ruixue

AU - Gong, Daoyuan

AU - He, Haijian

AU - Rock, Daniel L.

AU - Hao, Wenbo

AU - Luo, Shuhong

N1 - Funding Information: This study was supported by the National Natural Science Foundation of China [grant numbers: 81773271 and 31672536 ] This project was partially supported by the Foshan University senior talent start fund [grant number: 20161110004 ]; the Foshan University high - level university fund [grant number: 20170131020 ]; and the Agriculture and Food Research Initiative Competitive from the USDA National Institute of Food and Agriculture [grant number: 2012-67015-19289 ]. Funding Information: This study was supported by the National Natural Science Foundation of China [grant numbers: 81773271 and 31672536] This project was partially supported by the Foshan University senior talent start fund [grant number: 20161110004]; the Foshan University high - level university fund [grant number: 20170131020]; and the Agriculture and Food Research Initiative Competitive from the USDA National Institute of Food and Agriculture [grant number: 2012-67015-19289]. Publisher Copyright: © 2018

PY - 2018/6/30

Y1 - 2018/6/30

N2 - Objective: Orf virus (ORFV) is the pathogen causing contagious pustular dermatitis in goats, sheep and herdsmen. Evidence has confirmed that ORFV can be used as a preventive and therapeutic immunomodulatory agent in several animal models. Our previous data demonstrated that ORFV024 is able to inhibit activation of the NF-κB signaling pathway and act as an important modulator for early immune responses against viral infection. However, the molecular mechanism by which ORFV024 exerting biological function remains unclear. In the present study, we explored and analyzed the function of host cellular proteins that interact with ORFV024. Methods: The yeast two-hybrid (Y2H) assay was performed to screen proteins interacting with ORFV024 using a cDNA library derived from primary ovine fetal turbinate cells (OFTu). Two of the screened proteins were further confirmed by confocal microscopy, His-tag pull-down assay and CO-Immunoprecipitation (CO-IP) assay. In addition, the ORFV024 interaction network was constructed using the STRING database. Results: In this study, 11 ovine cellular proteins were found to interact with ORFV024. In view of the importance of LAGE3 and IGFBP6 in the ORFV024 functional analysis, we further constructed LAGE3 and IGFBP6 interaction networks. The interactions between ORFV024 and LAGE3 or IGFBP6 were confirmed by confocal microscopy, LAGE3 was further confirmed in the His-tag pull-down assay and CO-IP assay. Conclusions: Our findings indicate that ORFV024 can interact with ovine cellular proteins LAGE3 and IGFBP6.

AB - Objective: Orf virus (ORFV) is the pathogen causing contagious pustular dermatitis in goats, sheep and herdsmen. Evidence has confirmed that ORFV can be used as a preventive and therapeutic immunomodulatory agent in several animal models. Our previous data demonstrated that ORFV024 is able to inhibit activation of the NF-κB signaling pathway and act as an important modulator for early immune responses against viral infection. However, the molecular mechanism by which ORFV024 exerting biological function remains unclear. In the present study, we explored and analyzed the function of host cellular proteins that interact with ORFV024. Methods: The yeast two-hybrid (Y2H) assay was performed to screen proteins interacting with ORFV024 using a cDNA library derived from primary ovine fetal turbinate cells (OFTu). Two of the screened proteins were further confirmed by confocal microscopy, His-tag pull-down assay and CO-Immunoprecipitation (CO-IP) assay. In addition, the ORFV024 interaction network was constructed using the STRING database. Results: In this study, 11 ovine cellular proteins were found to interact with ORFV024. In view of the importance of LAGE3 and IGFBP6 in the ORFV024 functional analysis, we further constructed LAGE3 and IGFBP6 interaction networks. The interactions between ORFV024 and LAGE3 or IGFBP6 were confirmed by confocal microscopy, LAGE3 was further confirmed in the His-tag pull-down assay and CO-IP assay. Conclusions: Our findings indicate that ORFV024 can interact with ovine cellular proteins LAGE3 and IGFBP6.

KW - IGFBP6

KW - LAGE3

KW - NF-κB signaling pathway

KW - ORFV024

KW - Orf virus

KW - Yeast two-hybrid system

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DO - 10.1016/j.gene.2018.03.089

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JO - Gene

JF - Gene

SN - 0378-1119

ER -

Identification of host cellular proteins LAGE3 and IGFBP6 that interact with orf virus protein ORFV024

Abstract

Keywords

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