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. 2019 Feb 5;31(12):867–875. doi: 10.3967/bes2018.115

Coxsackievirus B3 Infection Triggers Autophagy through 3 Pathways of Endoplasmic Reticulum Stress

Xiao Nuan LUO a,&, Hai Lan YAO b,&, Juan SONG a, Qin Qin SONG a, Bing Tian SHI a, Dong XIA a, Jun HAN a,#
PMCID: PMC7126911  PMID: 30636656

Abstract

Objective

Autophagy is a highly conserved intracellular degradation pathway. Many picornaviruses induce autophagy to benefit viral replication, but an understanding of how autophagy occurs remains incomplete. In this study, we explored whether coxsackievirus B3 (CVB3) infection induced autophagy through endoplasmic reticulum (ER) stress.

Methods

In CVB3-infected HeLa cells, the specific molecules of ER stress and autophagy were detected using Western blotting, reverse transcription polymerase chain reaction (RT-PCR), and confocal microscopy. Then PKR-like ER protein kinase (PERK) inhibitor, inositol-requiring protein-1 (IRE1) inhibitor, or activating transcription factor-6 (ATF6) inhibitor worked on CVB3-infected cells, their effect on autophagy was assessed by Western blotting for detecting microtubule-associated protein light chain 3 (LC3).

Results

CVB3 infection induced ER stress, and ER stress sensors PERK/eIF2α, IRE1/XBP1, and ATF6 were activated. CVB3 infection increased the accumulation of green fluorescent protein (GFP)-LC3 punctuation and induced the conversion from LC3-I to phosphatidylethanolamine-conjugated LC3-1 (LC3-II). CVB3 infection still decreased the expression of mammalian target of rapamycin (mTOR) and p-mTOR. Inhibition of PERK, IRE1, or ATF6 significantly decreased the ratio of LC3-II to LC3-I in CVB3-infected HeLa cells.

Conclusion

CVB3 infection induced autophagy through ER stress in HeLa cells, and PERK, IRE1, and ATF6a pathways participated in the regulation of autophagy. Our data suggested that ER stress may inhibit mTOR signaling pathway to induce autophagy during CVB3 infection.

Key words: Coxsackievirus B3 (CVB3), Autophagy, Endoplasmic reticulum (ER) stress, Unfolded protein response (UPR)

Biography

Biographical notes of first authors: LUO Xiao Nuan, female, born in 1989, PhD, majoring in immunology; YAO Hai Lan, female, born in 1975, PhD, majoring in immunology.

Footnotes

This study was supported by the China Mega-project for Infectious Disease [2018ZX10102001, 2018ZX10711001, 2018ZX10734401, and 2018ZX10734404]; and the SKLID Development Grant [2011SKLID104].

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