TY - JOUR
T1 - Interaction between chicken TRIM25 and MDA5 and their role in mediated antiviral activity against IBDV infection
AU - Diaz-Beneitez, Elisabet
AU - Cubas-Gaona, Liliana Lilibeth
AU - Candelas-Rivera, Oscar
AU - Benito-Zafra, Ana
AU - Sánchez-Aparicio, Maria Teresa
AU - Miorin, Lisa
AU - Rodríguez, José F.
AU - García-Sastre, Adolfo
AU - Rodríguez, Dolores
N1 - Funding Information:
This work was supported by grants AGL2017-87464-C2-1-P and PID2020-112847RB-I00 to DR and JR, funded by MCIN/AEI/10.13039/501100011033 and by “ERDF A way of making Europe,” by the European Union. This work was also partly supported by CRIPT (Center for Research on Influenza Pathogenesis and Transmission), a NIAID funded Center of Excellence for Influenza Research and Response (CEIRR, contract number 75N93021C00014) to AG-S. ED-B was supported by a FPU fellowship from the Spanish Ministry of Education, Culture and Sport. LC-G was recipient of a predoctoral research contract from the International Fellowship Program of the Caixa Foundation and an EMBO short-term fellowship (ASTF 245-2015). OC-R was supported by a contract from the Consejería de Juventud y Deporte de la Comunidad Autónoma de Madrid (PEJD-2017-PRE/BIO-5006). DR was recipient of a visiting scholar fellowship (PRX16/00538) sponsored by Fulbright Program and Spanish Ministry of Education, Culture and Sport. Additional funding was received from the Spanish National Research Council (CSIC) to support open access publication fees. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Funding Information:
We are grateful to Ana M. Oña from the Advanced Light Microscopy scientific CNB core facility her excellent technical assistance. We are also extremely grateful to Stephen Goodbourn (Institute for Infection and Immunity, St. George’s, University of London, United Kingdom) and José Manuel Martínez Costas (CIQUS. University of Santiago de Compostela. Spain) for generously sharing the plasmid expressing chMAVS and the avian reovirus, respectively.
Publisher Copyright:
Copyright © 2022 Diaz-Beneitez, Cubas-Gaona, Candelas-Rivera, Benito-Zafra, Sánchez-Aparicio, Miorin, Rodríguez, García-Sastre and Rodríguez.
PY - 2022/11/28
Y1 - 2022/11/28
N2 - Infectious Bursal Disease Virus (IBDV) is the causative agent of an immunosuppressive disease that affects domestic chickens (Gallus gallus) severely affecting poultry industry worldwide. IBDV infection is characterized by a rapid depletion of the bursal B cell population by apoptosis and the atrophy of this chief lymphoid organ. Previous results from our laboratory have shown that exposure of infected cells to type I IFN leads to an exacerbated apoptosis, indicating an important role of IFN in IBDV pathogenesis. It has been described that recognition of the dsRNA IBDV genome by MDA5, the only known cytoplasmic pattern recognition receptor for viral RNA in chickens, leads to type I IFN production. Here, we confirm that TRIM25, an E3 ubiquitin ligase that leads to RIG-I activation in mammalian cells, significantly contributes to positively regulate MDA5-mediated activation of the IFN-inducing pathway in chicken DF-1 cells. Ectopic expression of chTRIM25 together with chMDA5 or a deletion mutant version exclusively harboring the CARD domains (chMDA5 2CARD) enhances IFN-β and NF-ĸB promoter activation. Using co-immunoprecipitation assays, we show that chMDA5 interacts with chTRIM25 through the CARD domains. Moreover, chTRIM25 co-localizes with both chMDA5 and chMDA5 2CARD, but not with chMDA5 mutant proteins partially or totally lacking these domains. On the other hand, ablation of endogenous chTRIM25 expression reduces chMDA5-induced IFN-β and NF-ĸB promoter activation. Interestingly, ectopic expression of either wild-type chTRIM25, or a mutant version (chTRIM25 C59S/C62S) lacking the E3 ubiquitin ligase activity, restores the co-stimulatory effect of chMDA5 in chTRIM25 knockout cells, suggesting that the E3-ubiquitin ligase activity of chTRIM25 is not required for its downstream IFN-β and NF-ĸB activating function. Also, IBDV-induced expression of IFN-β, Mx and OAS genes was reduced in chTRIM25 knockout as compared to wild-type cells, hence contributing to the enhancement of IBDV replication. Enhanced permissiveness to replication of other viruses, such as avian reovirus, Newcastle disease virus and vesicular stomatitis virus was also observed in chTRIM25 knockout cells. Additionally, chTRIM25 knockout also results in reduced MAVS-induced IFN-β promoter stimulation. Nonetheless, similarly to its mammalian counterpart, chTRIM25 overexpression in wild-type DF-1 cells causes the degradation of ectopically expressed chMAVS.
AB - Infectious Bursal Disease Virus (IBDV) is the causative agent of an immunosuppressive disease that affects domestic chickens (Gallus gallus) severely affecting poultry industry worldwide. IBDV infection is characterized by a rapid depletion of the bursal B cell population by apoptosis and the atrophy of this chief lymphoid organ. Previous results from our laboratory have shown that exposure of infected cells to type I IFN leads to an exacerbated apoptosis, indicating an important role of IFN in IBDV pathogenesis. It has been described that recognition of the dsRNA IBDV genome by MDA5, the only known cytoplasmic pattern recognition receptor for viral RNA in chickens, leads to type I IFN production. Here, we confirm that TRIM25, an E3 ubiquitin ligase that leads to RIG-I activation in mammalian cells, significantly contributes to positively regulate MDA5-mediated activation of the IFN-inducing pathway in chicken DF-1 cells. Ectopic expression of chTRIM25 together with chMDA5 or a deletion mutant version exclusively harboring the CARD domains (chMDA5 2CARD) enhances IFN-β and NF-ĸB promoter activation. Using co-immunoprecipitation assays, we show that chMDA5 interacts with chTRIM25 through the CARD domains. Moreover, chTRIM25 co-localizes with both chMDA5 and chMDA5 2CARD, but not with chMDA5 mutant proteins partially or totally lacking these domains. On the other hand, ablation of endogenous chTRIM25 expression reduces chMDA5-induced IFN-β and NF-ĸB promoter activation. Interestingly, ectopic expression of either wild-type chTRIM25, or a mutant version (chTRIM25 C59S/C62S) lacking the E3 ubiquitin ligase activity, restores the co-stimulatory effect of chMDA5 in chTRIM25 knockout cells, suggesting that the E3-ubiquitin ligase activity of chTRIM25 is not required for its downstream IFN-β and NF-ĸB activating function. Also, IBDV-induced expression of IFN-β, Mx and OAS genes was reduced in chTRIM25 knockout as compared to wild-type cells, hence contributing to the enhancement of IBDV replication. Enhanced permissiveness to replication of other viruses, such as avian reovirus, Newcastle disease virus and vesicular stomatitis virus was also observed in chTRIM25 knockout cells. Additionally, chTRIM25 knockout also results in reduced MAVS-induced IFN-β promoter stimulation. Nonetheless, similarly to its mammalian counterpart, chTRIM25 overexpression in wild-type DF-1 cells causes the degradation of ectopically expressed chMAVS.
KW - IBDV
KW - MDA5
KW - TRIM25
KW - innate immunity
KW - interferon
UR - http://www.scopus.com/inward/record.url?scp=85143972981&partnerID=8YFLogxK
U2 - 10.3389/fmicb.2022.1068328
DO - 10.3389/fmicb.2022.1068328
M3 - Article
AN - SCOPUS:85143972981
SN - 1664-302X
VL - 13
JO - Frontiers in Microbiology
JF - Frontiers in Microbiology
M1 - 1068328
ER -