Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Mar;25(2):e21.
doi: 10.4142/jvs.23236.

Peste des petits ruminants virus infection induces endoplasmic reticulum stress and apoptosis via IRE1-XBP1 and IRE1-JNK signaling pathways

Shuyi Yuan  1 , Yanfen Liu  1 , Yun Mu  1 , Yongshen Kuang  1 , Shaohong Chen  2 , Yun-Tao Zhao  2 , You Liu  3
Affiliations

Peste des petits ruminants virus infection induces endoplasmic reticulum stress and apoptosis via IRE1-XBP1 and IRE1-JNK signaling pathways

Shuyi Yuan et al. J Vet Sci. 2024 Mar.

Abstract

Background: Peste des petits ruminants (PPR) is a contagious and fatal disease of sheep and goats. PPR virus (PPRV) infection induces endoplasmic reticulum (ER) stress-mediated unfolded protein response (UPR). The activation of UPR signaling pathways and their impact on apoptosis and virus replication remains controversial.

Objectives: To investigate the role of PPRV-induced ER stress and the IRE1-XBP1 and IRE1-JNK pathways and their impact on apoptosis and virus replication.

Methods: The cell viability and virus replication were assessed by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay, immunofluorescence assay, and Western blot. The expression of ER stress biomarker GRP78, IRE1, and its downstream molecules, PPRV-N protein, and apoptosis-related proteins was detected by Western blot and quantitative reverse transcription-polymerase chain reaction, respectively. 4-Phenylbutyric acid (4-PBA) and STF-083010 were respectively used to inhibit ER stress and IRE1 signaling pathway.

Results: The expression of GRP78, IRE1α, p-IRE1α, XBP1s, JNK, p-JNK, caspase-3, caspase-9, Bax and PPRV-N were significantly up-regulated in PPRV-infected cells, the expression of Bcl-2 was significantly down-regulated. Due to 4-PBA treatment, the expression of GRP78, p-IRE1α, XBP1s, p-JNK, caspase-3, caspase-9, Bax, and PPRV-N were significantly down-regulated, the expression of Bcl-2 was significantly up-regulated. Moreover, in PPRV-infected cells, the expression of p-IRE1α, p-JNK, Bax, and PPRV-N was significantly decreased, and the expression of Bcl-2 was increased in the presence of STF-083010.

Conclusions: PPRV infection induces ER stress and IRE1 activation, resulting in apoptosis and enhancement of virus replication through IRE1-XBP1s and IRE1-JNK pathways.

Keywords: IRE1-JNK pathway; IRE1-XBP1 pathway; Peste des petits ruminants virus; apoptosis; endoplasmic reticulum stress.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Fig. 1
Fig. 1. Effect of PPRV infection on the viability of LDG-2 cells. Data are presented as mean ± SD.
PPRV, peste des petits ruminants virus. ***p < 0.001 vs. the cell control.
Fig. 2
Fig. 2. Propagation of PPRV in LDG-2 cells at 12, 18, 24, 30, and 36 hpi. (A) PPRV replication and distribution detected by IFA (scale bars = 100 µm). (B) Expression of PPRV-N protein detected by Western blot. (C) Quantitative analysis of PPRV-N protein. (D) mRNA expression of PPRV-N protein detected by qRT-PCR. Data are presented as mean ± SD.
PPRV, peste des petits ruminants virus; IFA, immunofluorescence assay; N, nucleocapsid; qRT-PCR, quantitative reverse transcription-polymerase chain reaction; DAPI, 4′,6-diamidino-2-phenylindole; FITC, fluorescein isothiocyanate. *p < 0.05, **p < 0.01, ***p < 0.001 vs. the cell control.
Fig. 3
Fig. 3. ER stress induced by PPRV infection. (A) GRP78 expression at 12, 18, 24, 30, and 36 hpi was detected by Western blot. (B) Quantitative analysis of GRP78. Data was presented as mean ± SD.
ER, endoplasmic reticulum; PPRV, peste des petits ruminants virus. **p < 0.01; ***p < 0.001 vs. the cell control.
Fig. 4
Fig. 4. Expression levels of proteins related to IRE1-XBP1s and IRE1-JNK signaling pathways in PPRV-infected cells. (A) Expression of p-IRE1α, IRE1α, XBP1s, p-JNK, and JNK detected by Western blot. (B) Ratio of p-IRE1α/IRE1α. (C) Quantitative analysis of XBP1s. (D) Ratios of p-JNK/JNK. Data are presented as mean ± SD.
PPRV, peste des petits ruminants virus. *p < 0.05, **p < 0.01, ***p < 0.001 vs. the cell control.
Fig. 5
Fig. 5. Effect of 4-PBA treatment on IRE1-XBP1s and IRE1-JNK signaling pathways in PPRV-infected cells. (A) Expression of GRP78, p-IRE1α, IRE1α, XBP1s, p-JNK, and JNK detected by Western blot in the absence or presence of 4-PBA; (B) Ratio of GRP78. (C) Ratio of p-IRE1α/IRE1α. (D) Quantitative analysis of XBP1s. (E) Ratios of p-JNK/JNK. Data are presented as mean ± SD.
4-PBA, 4-phenylbutyric acid; PPRV, peste des petits ruminants virus. *p < 0.05, **p < 0.01, ***p < 0.001 vs. the virus control.
Fig. 6
Fig. 6. Effect of 4-PBA treatment on PPRV-N protein expression in LDG-2 cells. (A) PPRV-N protein by Western blot in the absence or presence of 4-PBA. (B) Quantitative analysis of PPRV-N protein. Data are presented as mean ± SD.
4-PBA, 4-phenylbutyric acid; PPRV, peste des petits ruminants virus; N, nucleocapsid. ***p < 0.001 vs. the virus control.
Fig. 7
Fig. 7. Effect of PPRV infection on Bcl-2 and Bax, caspase-3, and Ccaspase-9. (A)Expression of Bcl-2 and Bax detected by Western blot. (B) Ratios of Bcl-2/Bax. (C) Expression of Bcl-2 and Bax detected by Western blot in the absence or presence of 4-PBA. (D) Ratios of Bcl-2/Bax in the absence or presence of 4-PBA. (E) mRNA expression of caspase-3. (F) mRNA expression of caspase-9. (G) mRNA expression of caspase-3 in the absence or presence of 4-PBA. (H) mRNA expression of caspase-9 in the absence or presence of 4-PBA. Data are presented as mean ± SD.
PPRV, peste des petits ruminants virus; 4-PBA, 4-phenylbutyric acid. *p < 0.05, **p < 0.01, ***p < 0.001 vs. the cell control; ##p < 0.01, ###p < 0.001 vs. the virus control.
Fig. 8
Fig. 8. PPRV induces apoptosis through the IRE1-JNK pathway to promote viral replication. (A) Expression of p-IRE1α, IRE1α, XBP1s, p-JNK, JNK, Bcl-2, Bax, and PPRV-N detected by Western blot in the absence or presence of STF-083010. (B) Ratios of p-IRE1α/IRE1α; (C) Ratio of p-JNK/JNK. (D) Quantitative analysis of PPRV-N protein expression. (E) Ratios of Bcl-2/Bax. Data are presented as mean ± SD.
PPRV, peste des petits ruminants virus; N, nucleocapsid. **p < 0.01, ***p < 0.001 vs. the virus control.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Kumar N, Maherchandani S, Kashyap SK, Singh SV, Sharma S, Chaubey KK, et al. Peste des petits ruminants virus infection of small ruminants: a comprehensive review. Viruses. 2014;6(6):2287–2327. - PMC - PubMed
    1. Salminen A, Kaarniranta K, Kauppinen A. ER stress activates immunosuppressive network: implications for aging and Alzheimer’s disease. J Mol Med (Berl) 2020;98(5):633–650. - PMC - PubMed
    1. Liu Y, Wang M, Cheng A, Yang Q, Wu Y, Jia R, et al. The role of host eIF2α in viral infection. Virol J. 2020;17(1):112. - PMC - PubMed
    1. Cheng JH, Sun YJ, Zhang FQ, Zhang XR, Qiu XS, Yu LP, et al. Newcastle disease virus NP and P proteins induce autophagy via the endoplasmic reticulum stress-related unfolded protein response. Sci Rep. 2016;6(1):24721. - PMC - PubMed
    1. Wang W, Bi Z, Liu Y, Xia X, Qian J, Tan Y, et al. The H protein of attenuated canine distemper virus is degraded via endoplasmic reticulum-associated protein degradation. Front Vet Sci. 2023;10:1214318. - PMC - PubMed