Nuclear ribonucleoprotein RALY targets virus nucleocapsid protein and induces autophagy to restrict porcine epidemic diarrhea virus replication

Abstract

Porcine epidemic diarrhea virus (PEDV) causes diarrhea and dehydration in pigs and leads to great economic losses in the commercial swine industry. However, the underlying molecular mechanisms of host response to viral infection remain unclear. In the present study, we investigated a novel mechanism by which RALY, a member of the heterogeneous nuclear ribonucleoprotein family, significantly promotes the degradation of the PEDV nucleocapsid (N) protein to inhibit viral replication. Furthermore, we identified an interaction between RALY and the E3 ubiquitin ligase MARCH8 (membrane-associated RING-CH 8), as well as the cargo receptor NDP52 (nuclear dot protein 52 kDa), suggesting that RALY could suppress PEDV replication by degrading the viral N protein through a RALY–MARCH8–NDP52–autophagosome pathway. Collectively, these results suggest a preventive role of RALY against PEDV infection via the autophagy pathway and open up the possibility of inducing RALY in vivo as an effective prophylactic and preventive treatment for PEDV infection. Porcine epidemic diarrhea virus (PEDV) causes diarrhea and dehydration in pigs and leads to great economic losses in the commercial swine industry. However, the underlying molecular mechanisms of host response to viral infection remain unclear. In the present study, we investigated a novel mechanism by which RALY, a member of the heterogeneous nuclear ribonucleoprotein family, significantly promotes the degradation of the PEDV nucleocapsid (N) protein to inhibit viral replication. Furthermore, we identified an interaction between RALY and the E3 ubiquitin ligase MARCH8 (membrane-associated RING-CH 8), as well as the cargo receptor NDP52 (nuclear dot protein 52 kDa), suggesting that RALY could suppress PEDV replication by degrading the viral N protein through a RALY–MARCH8–NDP52–autophagosome pathway. Collectively, these results suggest a preventive role of RALY against PEDV infection via the autophagy pathway and open up the possibility of inducing RALY in vivo as an effective prophylactic and preventive treatment for PEDV infection. Porcine epidemic diarrhea virus (PEDV), a member of the genus Alphacoronavirus in the family Coronaviridae, causes acute diarrhea, vomiting, dehydration, and high mortality in neonatal piglets (1Jung K. Saif L.J. Wang Q. Porcine epidemic diarrhea virus (PEDV): an update on etiology, transmission, pathogenesis, and prevention and control.Virus Res. 2020; 286: 198045Crossref PEDV a of the for and nucleocapsid and an protein K. of the epidemic (PEDV) epidemic diarrhea economic losses to the swine the the to the an on the of Res. on prophylactic and treatment to PEDV infection as a member of the heterogeneous nuclear ribonucleoprotein family of that an role in and of the protein the in Res. of RALY Furthermore, RALY to in L.J. of the of a novel L.J. molecular for the and of the to by the of the Wang Wang by the of RALY in the and the of protein RALY promotes protein and in Wang an and a in Res. 2020; of RALY promotes and in Res. and RALY and by the of the a that and for as in autophagy in and as an mechanism in and in and K. autophagy virus infection and to autophagy epidemic diarrhea virus in Res. PEDV N protein an role in the to viral replication Q. of and viral of Res. N protein in the PEDV replication and a to inhibit virus replication by that host as response epidemic diarrhea virus replication by to viral nucleocapsid protein epidemic diarrhea virus replication by to viral nucleocapsid protein Wang replication by degrading nucleocapsid protein through and epidemic diarrhea virus by degradation of the nucleocapsid suppress PEDV replication by and degrading the viral N protein via to the that the of RALY to of the present study, we to the host mechanism in PEDV infection by the molecular mechanisms by RALY PEDV infection. that RALY PEDV replication by the degradation pathway. RALY as a in PEDV infection between RALY and the of PEDV infection on RALY we the and protein of RALY viral infection. PEDV a of infection of as Q. Wang of epidemic diarrhea virus by the and the of RALY by and and infection results that protein and of RALY significantly in in and that PEDV infection inhibit RALY in host to and as an protein protein RALY via in and an role in and Wang an and a in Res. 2020; the of RALY in PEDV PEDV an of and that PEDV N protein and significantly and in and RALY the replication of the virus RALY and results suggest that RALY suppress PEDV replication in PEDV replication in the of RALY on PEDV an the PEDV an of and and and for PEDV N and viral in viral and and the results that the of RALY significantly PEDV replication. 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PEDV RALY E3 ubiquitin ligase MARCH8 to ubiquitin viral N protein and the cargo receptor NDP52 to the N protein and to for RING-CH nucleocapsid nuclear dot protein 52 epidemic diarrhea a member of the heterogeneous nuclear ribonucleoprotein family, RALY protein identified as a which could the nuclear a viral protein by the virus an that of the virus and which a heterogeneous nuclear RALY an the N nuclear and a the of the protein the in Res. which to the of RALY and by the of the and in and role in and the of RALY and a for protein RALY via in we that PEDV infection RALY in host and of RALY PEDV replication infection in and Furthermore, we that RALY the N and RALY in the to the the N results that RALY a for PEDV a the of which and cargo protein and to for as host autophagy and cargo in K. the autophagy 2020; of the autophagy that the PEDV to autophagy epidemic diarrhea virus replication by to viral nucleocapsid epidemic diarrhea virus by degradation of the nucleocapsid Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; In study, we that the degradation of N protein by RALY by autophagy and to the role of autophagy in PEDV infection autophagy epidemic diarrhea virus in Res. we that RALY a against PEDV infection by the degradation of viral the PEDV a we host E3 ubiquitin ligase MARCH8 to the of PEDV N inducing degradation Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; MARCH8 a member of the E3 ubiquitin ligase family and to inhibit and virus infection infection by of K. MARCH8 viral infection by 2020; K. the that to that MARCH8 the of MARCH8 the PEDV N and the PEDV N protein by in degradation of the N protein through the autophagy pathway to suppress viral replication in Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; K. the that to by the ligase of inducing receptor we that of MARCH8 NDP52 inhibit the degradation of the PEDV N Furthermore, RALY–MARCH8–NDP52–autophagosome pathway a role in N protein degradation and the host against PEDV we that RALY PEDV replication by degrading the viral N PEDV RALY the E3 ubiquitin ligase MARCH8 to the of the PEDV N the cargo receptor NDP52 the ubiquitin and N protein to for we for a novel mechanism of viral infection role of RALY in the host PEDV infection the for a and in in by and in PEDV and in and and and NDP52 and MARCH8 and and against the PEDV N protein in and viral in the to the PEDV an of for by in for PEDV the and in a and for in by and to the and and the by and the to the and to the to the in for and in a by the and in by the and RALY and in to protein to the and by for and for for and the for a for in the as Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; for a and for results of the Porcine epidemic diarrhea virus (PEDV), a member of the genus Alphacoronavirus in the family Coronaviridae, causes acute diarrhea, vomiting, dehydration, and high mortality in neonatal piglets (1Jung K. Saif L.J. Wang Q. Porcine epidemic diarrhea virus (PEDV): an update on etiology, transmission, pathogenesis, and prevention and control.Virus Res. 2020; 286: 198045Crossref PEDV a of the for and nucleocapsid and an protein K. of the epidemic (PEDV) epidemic diarrhea economic losses to the swine the the to the an on the of Res. on prophylactic and treatment to PEDV infection RALY as a member of the heterogeneous nuclear ribonucleoprotein family of that an role in and of the protein the in Res. of RALY Furthermore, RALY to in L.J. of the of a novel L.J. molecular for the and of the to by the of the Wang Wang by the of RALY in the and the of protein RALY promotes protein and in Wang an and a in Res. 2020; of RALY promotes and in Res. and RALY and by the of the a that and for as in autophagy in and as an mechanism in and in and K. autophagy virus infection and to autophagy epidemic diarrhea virus in Res. PEDV N protein an role in the to viral replication Q. of and viral of Res. N protein in the PEDV replication and a to inhibit virus replication by that host as response epidemic diarrhea virus replication by to viral nucleocapsid protein epidemic diarrhea virus replication by to viral nucleocapsid protein Wang replication by degrading nucleocapsid protein through and epidemic diarrhea virus by degradation of the nucleocapsid suppress PEDV replication by and degrading the viral N protein via to the that the of RALY to of In the present study, we to the host mechanism in PEDV infection by the molecular mechanisms by RALY PEDV infection. that RALY PEDV replication by the degradation pathway. RALY as a in PEDV infection between RALY and the of PEDV infection on RALY we the and protein of RALY viral infection. PEDV a of infection of as Q. Wang of epidemic diarrhea virus by the and the of RALY by and and infection results that protein and of RALY significantly in in and that PEDV infection inhibit RALY in host to and as an protein protein RALY via in and an role in and Wang an and a in Res. 2020; the of RALY in PEDV PEDV an of and that PEDV N protein and significantly and in and RALY the replication of the virus RALY and results suggest that RALY suppress PEDV replication in PEDV replication in the of RALY on PEDV an the PEDV an of and and and for PEDV N and viral in viral and and the results that the of RALY significantly PEDV replication. Furthermore, the on viral replication RALY as the N protein the of and viral in the of the that the of RALY PEDV replication that RALY PEDV replication in PEDV N protein N via the mechanism by RALY to PEDV we the interaction of RALY PEDV and for PEDV protein and PEDV N protein to by and the interaction by the of in the that PEDV N protein RALY, the of In a which between PEDV N and RALY Furthermore, and and protein by RALY in the and to the to N in the In these that RALY the N protein of PEDV N protein N by and for by by interaction of PEDV N protein the RALY and PEDV N and and in for the by and for and and and the of and N protein by and and and by nucleocapsid epidemic diarrhea N protein in the viral replication and pathogenesis, as the host and nucleocapsid a the mechanism by which RALY the PEDV N we and the of N we that RALY N protein in a the the autophagy and results the degradation of N protein by the autophagy and Collectively, these results that RALY promotes the degradation of PEDV N protein through PEDV N protein through the RING-CH dot protein 52 study, we that autophagy by PEDV infection Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; host as and could E3 ubiquitin ligase MARCH8 (membrane-associated RING-CH to PEDV N protein and the N protein could by the cargo receptor NDP52 (nuclear dot protein 52 and to the for degradation Wang replication by degrading nucleocapsid protein through Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; the mechanism of degradation of PEDV N via we a and that the E3 ubiquitin ligase and which by and the of RALY to MARCH8 and in the and PEDV N protein through the RALY–MARCH8–NDP52–autophagosome pathway. and for by by interaction of MARCH8 the RALY and MARCH8 in for the by and and and and the as and by by interaction of MARCH8 the RALY and NDP52 in for the by and and and and the and the and the of the MARCH8 NDP52 by and MARCH8 NDP52 N protein by and MARCH8 NDP52 PEDV an of and for of PEDV N protein RING-CH of nucleocapsid nuclear dot protein 52 epidemic diarrhea we the between RALY and the cargo receptor of RALY which by by and that RALY the to the and NDP52 that the pathway for PEDV N protein and MARCH8 NDP52 and that of MARCH8 NDP52 degradation of PEDV N protein we MARCH8 NDP52 the PEDV an of results that of MARCH8 NDP52 the of PEDV infection results that RALY PEDV N protein through the RALY–MARCH8–NDP52–autophagosome pathway. between RALY and the of PEDV infection on RALY we the and protein of RALY viral infection. 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In study, we that autophagy by PEDV infection Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; host as and could E3 ubiquitin ligase MARCH8 (membrane-associated RING-CH to PEDV N protein and the N protein could by the cargo receptor NDP52 (nuclear dot protein 52 and to the for degradation Wang replication by degrading nucleocapsid protein through Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; the mechanism of degradation of PEDV N via we a and that the E3 ubiquitin ligase and which by and the of RALY to MARCH8 and in the and we the between RALY and the cargo receptor of RALY which by by and that RALY the to the and NDP52 that the pathway for PEDV N protein and MARCH8 NDP52 and that of MARCH8 NDP52 degradation of PEDV N protein we MARCH8 NDP52 the PEDV an of results that of MARCH8 NDP52 the of PEDV infection results that RALY PEDV N protein through the RALY–MARCH8–NDP52–autophagosome pathway. causes diarrhea and vomiting, and of of a to swine In study, we investigated the host PEDV infection and viral replication. identified RALY as a to suppress PEDV replication by degrading the viral N protein through the RALY–MARCH8–NDP52–autophagosome pathway a member of the heterogeneous nuclear ribonucleoprotein family, RALY protein identified as a which could the nuclear a viral protein by the virus an that of the virus and which a heterogeneous nuclear RALY an the N nuclear and a the of the protein the in Res. which to the of RALY and by the of the and in and role in and the of RALY and a for protein RALY via in we that PEDV infection RALY in host and of RALY PEDV replication infection in and Furthermore, we that RALY the N and RALY in the to the the N results that RALY a for PEDV a the of which and cargo protein and to for as host autophagy and cargo in K. the autophagy 2020; of the autophagy that the PEDV to autophagy epidemic diarrhea virus replication by to viral nucleocapsid epidemic diarrhea virus by degradation of the nucleocapsid Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; In study, we that the degradation of N protein by RALY by autophagy and to the role of autophagy in PEDV infection autophagy epidemic diarrhea virus in Res. we that RALY a against PEDV infection by the degradation of viral the PEDV a we host E3 ubiquitin ligase MARCH8 to the of PEDV N inducing degradation Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; MARCH8 a member of the E3 ubiquitin ligase family and to inhibit and virus infection infection by of K. MARCH8 viral infection by 2020; K. the that to that MARCH8 the of MARCH8 the PEDV N and the PEDV N protein by in degradation of the N protein through the autophagy pathway to suppress viral replication in Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; K. the that to by the ligase of inducing receptor we that of MARCH8 NDP52 inhibit the degradation of the PEDV N Furthermore, RALY–MARCH8–NDP52–autophagosome pathway a role in N protein degradation and the host against PEDV we that RALY PEDV replication by degrading the viral N PEDV RALY the E3 ubiquitin ligase MARCH8 to the of the PEDV N the cargo receptor NDP52 the ubiquitin and N protein to for we for a novel mechanism of viral infection role of RALY in the host PEDV infection the for a PEDV causes diarrhea and vomiting, and of of a to swine In study, we investigated the host PEDV infection and viral replication. identified RALY as a to suppress PEDV replication by degrading the viral N protein through the RALY–MARCH8–NDP52–autophagosome pathway a member of the heterogeneous nuclear ribonucleoprotein family, RALY protein identified as a which could the nuclear a viral protein by the virus an that of the virus and which a heterogeneous nuclear RALY an the N nuclear and a the of the protein the in Res. which to the of RALY and by the of the and in and role in and the of RALY and a for protein RALY via in we that PEDV infection RALY in host and of RALY PEDV replication infection in and Furthermore, we that RALY the N and RALY in the to the the N results that RALY a for PEDV infection. a the of which and cargo protein and to for as host autophagy and cargo in K. the autophagy 2020; of the autophagy that the PEDV to autophagy epidemic diarrhea virus replication by to viral nucleocapsid epidemic diarrhea virus by degradation of the nucleocapsid Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; In study, we that the degradation of N protein by RALY by autophagy and to the role of autophagy in PEDV infection autophagy epidemic diarrhea virus in Res. we that RALY a against PEDV infection by the degradation of viral the PEDV infection. In a we host E3 ubiquitin ligase MARCH8 to the of PEDV N inducing degradation Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; MARCH8 a member of the E3 ubiquitin ligase family and to inhibit and virus infection infection by of K. MARCH8 viral infection by 2020; K. the that to that MARCH8 the of MARCH8 the PEDV N and the PEDV N protein by in degradation of the N protein through the autophagy pathway to suppress viral replication in Wang epidemic diarrhea virus replication by and degrading virus nucleocapsid protein 2020; K. the that to by the ligase of inducing receptor we that of MARCH8 NDP52 inhibit the degradation of the PEDV N Furthermore, RALY–MARCH8–NDP52–autophagosome pathway a role in N protein degradation and the host against PEDV infection. 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