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Porcine epidemic diarrhea virus S1 protein is the critical inducer of apoptosis

Identifieur interne : 000C52 ( Main/Exploration ); précédent : 000C51; suivant : 000C53

Porcine epidemic diarrhea virus S1 protein is the critical inducer of apoptosis

Auteurs : Yifeng Chen [République populaire de Chine] ; Zhibang Zhang [République populaire de Chine] ; Jie Li [République populaire de Chine] ; Yueyi Gao [République populaire de Chine] ; Lei Zhou [République populaire de Chine] ; Xinna Ge [République populaire de Chine] ; Jun Han [République populaire de Chine] ; Xin Guo [République populaire de Chine] ; Hanchun Yang [République populaire de Chine]

Source :

RBID : PMC:6222994

Descripteurs français

English descriptors

Abstract

Background

Porcine Epidemic Diarrhea (PED) is an acute and highly contagious enteric disease caused by PED virus (PEDV), characterized by vomitting, watery diarrhea and fatal dehydration with high mortality in sucking piglets of one week of age. Although PEDV induced cell apoptosis has been established in vitro and in vivo, the functional protein that contributes to this event remains unclear.

Methods

The activation or cleavage of main apoptosis-associated molecular such as AIFM1, caspase-3, caspase-8, caspase-9 and PARP in PEDV infected host cells were analyzed by western blotting. The nuclear change of infected cell was monitored by confocal immunofluorescence assay. The overexpressing plasmids of 16 non-structural proteins (Nsp1–16) and 6 structural proteins (M, N, E, ORF3, S1 and S2) were constructed by cloning. Cell apoptosis induced by PEDV or overexpression non-structural or structural proteins was measured by the flow cytometry assay.

Results

PEDV could infect various host cells including Vero, Vero-E6 and Marc-145 and cause obvious cytopathic effects, including roundup, cell fusion, cell membrane vacuolation, syncytium formation and cause apparent apoptosis. In infected cells, PEDV-induced apoptosis is accompanied by nuclear concentration and fragmentation as a result of caspase-3 and caspase-8 activation and AIFM1 and PARP cleavage. Overexpression of S1 Spike protein of PEDV SM98 strain effectively induced host cell apoptosis, while the expression of the other non-structure proteins (Nsp1–16) and structural proteins (M, N, E, S2 and ORF3) has no or less effect on cell apoptosis. Similarly, expression of S1 protein from wild-type strain BJ2011 or cell-adapted strain CV777, also induce apoptosis in transfected cells. Finally, we demonstrated that the S1 proteins from various coronavirus family members such as TGEV, IBV, CCoV, SARS and MERS could also induce Vero-E6 cells apoptosis.

Conclusion

S1 Spike protein is one of the most critical functional proteins that contribute to cell apoptosis. Expression of S1 proteins of the coronavirus tested in this study could all induce cell apoptosis suggesting S1 maybe is an effective inducer in Coronavirus-induced cell apoptosis and targeting S1 protein expression probably is a promising strategy to inhibit coronavirus infection and thus mediated apoptosis on host cells.

Electronic supplementary material

The online version of this article (10.1186/s12985-018-1078-4) contains supplementary material, which is available to authorized users.


Url:
DOI: 10.1186/s12985-018-1078-4
PubMed: 30404647
PubMed Central: 6222994


Affiliations:


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<term>Caspases (metabolism)</term>
<term>Chlorocebus aethiops</term>
<term>Coronavirus Infections (virology)</term>
<term>Cytopathogenic Effect, Viral</term>
<term>Poly(ADP-ribose) Polymerases (metabolism)</term>
<term>Porcine epidemic diarrhea virus (genetics)</term>
<term>Porcine epidemic diarrhea virus (pathogenicity)</term>
<term>Spike Glycoprotein, Coronavirus (genetics)</term>
<term>Spike Glycoprotein, Coronavirus (metabolism)</term>
<term>Swine</term>
<term>Swine Diseases (virology)</term>
<term>Vero Cells</term>
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<term>Animaux</term>
<term>Apoptose</term>
<term>Caspases (métabolisme)</term>
<term>Cellules Vero</term>
<term>Effet cytopathogène viral</term>
<term>Facteur inducteur d'apoptose (métabolisme)</term>
<term>Glycoprotéine de spicule des coronavirus (génétique)</term>
<term>Glycoprotéine de spicule des coronavirus (métabolisme)</term>
<term>Infections à coronavirus (virologie)</term>
<term>Maladies des porcs (virologie)</term>
<term>Poly(ADP-ribose) polymerases (métabolisme)</term>
<term>Suidae</term>
<term>Virus de la diarrhée porcine épidémique (génétique)</term>
<term>Virus de la diarrhée porcine épidémique (pathogénicité)</term>
</keywords>
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<term>Spike Glycoprotein, Coronavirus</term>
</keywords>
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<term>Apoptosis Inducing Factor</term>
<term>Caspases</term>
<term>Poly(ADP-ribose) Polymerases</term>
<term>Spike Glycoprotein, Coronavirus</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en">
<term>Porcine epidemic diarrhea virus</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr">
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Virus de la diarrhée porcine épidémique</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr">
<term>Caspases</term>
<term>Facteur inducteur d'apoptose</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Poly(ADP-ribose) polymerases</term>
</keywords>
<keywords scheme="MESH" qualifier="pathogenicity" xml:lang="en">
<term>Porcine epidemic diarrhea virus</term>
</keywords>
<keywords scheme="MESH" qualifier="pathogénicité" xml:lang="fr">
<term>Virus de la diarrhée porcine épidémique</term>
</keywords>
<keywords scheme="MESH" qualifier="virologie" xml:lang="fr">
<term>Infections à coronavirus</term>
<term>Maladies des porcs</term>
</keywords>
<keywords scheme="MESH" qualifier="virology" xml:lang="en">
<term>Coronavirus Infections</term>
<term>Swine Diseases</term>
</keywords>
<keywords scheme="MESH" xml:lang="en">
<term>Animals</term>
<term>Apoptosis</term>
<term>Chlorocebus aethiops</term>
<term>Cytopathogenic Effect, Viral</term>
<term>Swine</term>
<term>Vero Cells</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr">
<term>Animaux</term>
<term>Apoptose</term>
<term>Cellules Vero</term>
<term>Effet cytopathogène viral</term>
<term>Suidae</term>
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</teiHeader>
<front>
<div type="abstract" xml:lang="en">
<sec>
<title>Background</title>
<p id="Par1">Porcine Epidemic Diarrhea (PED) is an acute and highly contagious enteric disease caused by PED virus (PEDV), characterized by vomitting, watery diarrhea and fatal dehydration with high mortality in sucking piglets of one week of age. Although PEDV induced cell apoptosis has been established in vitro and in vivo, the functional protein that contributes to this event remains unclear.</p>
</sec>
<sec>
<title>Methods</title>
<p id="Par2">The activation or cleavage of main apoptosis-associated molecular such as AIFM1, caspase-3, caspase-8, caspase-9 and PARP in PEDV infected host cells were analyzed by western blotting. The nuclear change of infected cell was monitored by confocal immunofluorescence assay. The overexpressing plasmids of 16 non-structural proteins (Nsp1–16) and 6 structural proteins (M, N, E, ORF3, S1 and S2) were constructed by cloning. Cell apoptosis induced by PEDV or overexpression non-structural or structural proteins was measured by the flow cytometry assay.</p>
</sec>
<sec>
<title>Results</title>
<p id="Par3">PEDV could infect various host cells including Vero, Vero-E6 and Marc-145 and cause obvious cytopathic effects, including roundup, cell fusion, cell membrane vacuolation, syncytium formation and cause apparent apoptosis. In infected cells, PEDV-induced apoptosis is accompanied by nuclear concentration and fragmentation as a result of caspase-3 and caspase-8 activation and AIFM1 and PARP cleavage. Overexpression of S1 Spike protein of PEDV SM98 strain effectively induced host cell apoptosis, while the expression of the other non-structure proteins (Nsp1–16) and structural proteins (M, N, E, S2 and ORF3) has no or less effect on cell apoptosis. Similarly, expression of S1 protein from wild-type strain BJ2011 or cell-adapted strain CV777, also induce apoptosis in transfected cells. Finally, we demonstrated that the S1 proteins from various coronavirus family members such as TGEV, IBV, CCoV, SARS and MERS could also induce Vero-E6 cells apoptosis.</p>
</sec>
<sec>
<title>Conclusion</title>
<p id="Par4">S1 Spike protein is one of the most critical functional proteins that contribute to cell apoptosis. Expression of S1 proteins of the coronavirus tested in this study could all induce cell apoptosis suggesting S1 maybe is an effective inducer in Coronavirus-induced cell apoptosis and targeting S1 protein expression probably is a promising strategy to inhibit coronavirus infection and thus mediated apoptosis on host cells.</p>
</sec>
<sec>
<title>Electronic supplementary material</title>
<p>The online version of this article (10.1186/s12985-018-1078-4) contains supplementary material, which is available to authorized users.</p>
</sec>
</div>
</front>
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<author>
<name sortKey="Shi, D" uniqKey="Shi D">D Shi</name>
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<author>
<name sortKey="Feng, L" uniqKey="Feng L">L Feng</name>
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<name sortKey="Chan, Cm" uniqKey="Chan C">CM Chan</name>
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<author>
<name sortKey="Ma, Cw" uniqKey="Ma C">CW Ma</name>
</author>
<author>
<name sortKey="Chan, Wy" uniqKey="Chan W">WY Chan</name>
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<author>
<name sortKey="Chan, Hy" uniqKey="Chan H">HY Chan</name>
</author>
</analytic>
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<name sortKey="Tsoi, H" uniqKey="Tsoi H">H Tsoi</name>
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<author>
<name sortKey="Li, L" uniqKey="Li L">L Li</name>
</author>
<author>
<name sortKey="Chen, Zs" uniqKey="Chen Z">ZS Chen</name>
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<author>
<name sortKey="Lau, Kf" uniqKey="Lau K">KF Lau</name>
</author>
<author>
<name sortKey="Tsui, Sk" uniqKey="Tsui S">SK Tsui</name>
</author>
<author>
<name sortKey="Chan, Hy" uniqKey="Chan H">HY Chan</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
<author>
<name sortKey="Lin, Cw" uniqKey="Lin C">CW Lin</name>
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<author>
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</author>
<author>
<name sortKey="Hsieh, Th" uniqKey="Hsieh T">TH Hsieh</name>
</author>
<author>
<name sortKey="Shiu, Sy" uniqKey="Shiu S">SY Shiu</name>
</author>
<author>
<name sortKey="Li, Jy" uniqKey="Li J">JY Li</name>
</author>
</analytic>
</biblStruct>
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<name sortKey="Ziebuhr, J" uniqKey="Ziebuhr J">J Ziebuhr</name>
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</analytic>
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<analytic>
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<name sortKey="Kamitani, W" uniqKey="Kamitani W">W Kamitani</name>
</author>
<author>
<name sortKey="Narayanan, K" uniqKey="Narayanan K">K Narayanan</name>
</author>
<author>
<name sortKey="Huang, C" uniqKey="Huang C">C Huang</name>
</author>
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<name sortKey="Ikegami, T" uniqKey="Ikegami T">T Ikegami</name>
</author>
<author>
<name sortKey="Ito, N" uniqKey="Ito N">N Ito</name>
</author>
</analytic>
</biblStruct>
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<biblStruct>
<analytic>
<author>
<name sortKey="Reguera, J" uniqKey="Reguera J">J Reguera</name>
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<author>
<name sortKey="Mudgal, G" uniqKey="Mudgal G">G Mudgal</name>
</author>
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<name sortKey="Santiago, C" uniqKey="Santiago C">C Santiago</name>
</author>
<author>
<name sortKey="Casasnovas, Jm" uniqKey="Casasnovas J">JM Casasnovas</name>
</author>
</analytic>
</biblStruct>
<biblStruct>
<analytic>
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<name sortKey="Zhao, G" uniqKey="Zhao G">G Zhao</name>
</author>
<author>
<name sortKey="Shi, Sq" uniqKey="Shi S">SQ Shi</name>
</author>
<author>
<name sortKey="Yang, Y" uniqKey="Yang Y">Y Yang</name>
</author>
<author>
<name sortKey="Peng, Jp" uniqKey="Peng J">JP Peng</name>
</author>
</analytic>
</biblStruct>
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</div1>
</back>
</TEI>
<affiliations>
<list>
<country>
<li>République populaire de Chine</li>
</country>
<settlement>
<li>Pékin</li>
</settlement>
</list>
<tree>
<country name="République populaire de Chine">
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<name sortKey="Chen, Yifeng" sort="Chen, Yifeng" uniqKey="Chen Y" first="Yifeng" last="Chen">Yifeng Chen</name>
</noRegion>
<name sortKey="Chen, Yifeng" sort="Chen, Yifeng" uniqKey="Chen Y" first="Yifeng" last="Chen">Yifeng Chen</name>
<name sortKey="Gao, Yueyi" sort="Gao, Yueyi" uniqKey="Gao Y" first="Yueyi" last="Gao">Yueyi Gao</name>
<name sortKey="Ge, Xinna" sort="Ge, Xinna" uniqKey="Ge X" first="Xinna" last="Ge">Xinna Ge</name>
<name sortKey="Guo, Xin" sort="Guo, Xin" uniqKey="Guo X" first="Xin" last="Guo">Xin Guo</name>
<name sortKey="Han, Jun" sort="Han, Jun" uniqKey="Han J" first="Jun" last="Han">Jun Han</name>
<name sortKey="Li, Jie" sort="Li, Jie" uniqKey="Li J" first="Jie" last="Li">Jie Li</name>
<name sortKey="Yang, Hanchun" sort="Yang, Hanchun" uniqKey="Yang H" first="Hanchun" last="Yang">Hanchun Yang</name>
<name sortKey="Zhang, Zhibang" sort="Zhang, Zhibang" uniqKey="Zhang Z" first="Zhibang" last="Zhang">Zhibang Zhang</name>
<name sortKey="Zhou, Lei" sort="Zhou, Lei" uniqKey="Zhou L" first="Lei" last="Zhou">Lei Zhou</name>
</country>
</tree>
</affiliations>
</record>

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