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      Porcine Epidemic Diarrhea Virus Induces Autophagy to Benefit Its Replication

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          Abstract

          The new porcine epidemic diarrhea (PED) has caused devastating economic losses to the swine industry worldwide. Despite extensive research on the relationship between autophagy and virus infection, the concrete role of autophagy in porcine epidemic diarrhea virus (PEDV) infection has not been reported. In this study, autophagy was demonstrated to be triggered by the effective replication of PEDV through transmission electron microscopy, confocal microscopy, and Western blot analysis. Moreover, autophagy was confirmed to benefit PEDV replication by using autophagy regulators and RNA interference. Furthermore, autophagy might be associated with the expression of inflammatory cytokines and have a positive feedback loop with the NF-κB signaling pathway during PEDV infection. This work is the first attempt to explore the complex interplay between autophagy and PEDV infection. Our findings might accelerate our understanding of the pathogenesis of PEDV infection and provide new insights into the development of effective therapeutic strategies.

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          Guidelines for the use and interpretation of assays for monitoring autophagy.

          C N Estevez, S M Lee, Qing Zhong … (2012)
          In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.
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            Porcine epidemic diarrhoea virus: a comprehensive review of molecular epidemiology, diagnosis, and vaccines

            Daesub Song, Bongkyun Park (2012)
            The porcine epidemic diarrhoea virus (PEDV), a member of the Coronaviridae family, causes acute diarrhoea and dehydration in pigs. Although it was first identified in Europe, it has become increasingly problematic in many Asian countries, including Korea, China, Japan, the Philippines, and Thailand. The economic impacts of the PEDV are substantial, given that it results in significant morbidity and mortality in neonatal piglets and is associated with increased costs related to vaccination and disinfection. Recently, progress has been made in understanding the molecular epidemiology of PEDV, thereby leading to the development of new vaccines. In the current review, we first describe the molecular and genetic characteristics of the PEDV. Then we discuss its molecular epidemiology and diagnosis, what vaccines are available, and how PEDV can be treated.
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              Coronaviruses Hijack the LC3-I-Positive EDEMosomes, ER-Derived Vesicles Exporting Short-Lived ERAD Regulators, for Replication

              Fulvio Reggiori, Iryna Monastyrska, Monique Verheije … (2010)
              Summary Coronaviruses (CoV), including SARS and mouse hepatitis virus (MHV), are enveloped RNA viruses that induce formation of double-membrane vesicles (DMVs) and target their replication and transcription complexes (RTCs) on the DMV-limiting membranes. The DMV biogenesis has been connected with the early secretory pathway. CoV-induced DMVs, however, lack conventional endoplasmic reticulum (ER) or Golgi protein markers, leaving their membrane origins in question. We show that MHV co-opts the host cell machinery for COPII-independent vesicular ER export of a short-living regulator of ER-associated degradation (ERAD), EDEM1, to derive cellular membranes for replication. MHV infection causes accumulation of EDEM1 and OS-9, another short-living ER chaperone, in the DMVs. DMVs are coated with the nonlipidated LC3/Atg8 autophagy marker. Downregulation of LC3, but not inactivation of host cell autophagy, protects cells from CoV infection. Our study identifies the host cellular pathway hijacked for supplying CoV replication membranes and describes an autophagy-independent role for nonlipidated LC3-I.
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                Author and article information

                Contributors
                Linda Dixon: Role: Academic Editor
                Simon Graham: Role: Academic Editor
                Journal
                Journal ID (nlm-ta): Viruses
                Journal ID (iso-abbrev): Viruses
                Journal ID (publisher-id): viruses
                Title: Viruses
                Publisher: MDPI
                ISSN (Electronic): 1999-4915
                Publication date (Electronic): 19 March 2017
                Publication date Collection: March 2017
                Volume: 9
                Issue: 3
                Electronic Location Identifier: 53
                Affiliations
                [1 ]State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; guoxiaozhen@ 123456webmail.hzau.edu.cn (X.G.); zhangmengjia@ 123456webmail.hzau.edu.cn (M.Z.); 632071039@ 123456webmail.hzau.edu.cn (X.Z.); wstx1992@ 123456163.com (X.T.); 15071194245@ 123456163.com (H.G.); aiyouwei0620@ 123456163.com (W.Z.); Memonatta80@ 123456webmail.hzau.edu.cn (A.M.M.); lzh1990@ 123456webmail.hzau.edu.cn (Z.L.); yingzizhu10@ 123456163.com (Y.Z.); abing0313@ 123456webmail.hzau.edu.cn (B.Z.); kuxugang84@ 123456163.com (X.K.); wumeizhou@ 123456mail.hzau.edu.cn (M.W.)
                [2 ]The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China; gkyangk@ 123456163.com
                [3 ]College of Animal Sciences and Technology, Huazhong Agricultural University, Wuhan 430070, China
                Author notes
                [* ]Correspondence: fanshengxian@ 123456mail.hzau.edu.cn (S.F.); he628@ 123456mail.hzau.edu.cn (Q.H.); Tel.: +86-27-8728-3005 (S.F.); +86-27-8728-6974 (Q.H.)
                Article
                Publisher ID: viruses-09-00053
                DOI: 10.3390/v9030053
                PMC ID: 5371808
                PubMed ID: 28335505
                SO-VID: 5ef25076-4b6d-4dac-8230-3d1697f49447
                Copyright © © 2017 by the authors.
                License:

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                Date received : 28 December 2016
                Date accepted : 15 March 2017
                Categories
                Subject: Article

                ScienceOpen disciplines: Microbiology & Virology
                Keywords: autophagy machinery,pedv replication,inflammatory responses,apoptosis
                Data availability:
                ScienceOpen disciplines: Microbiology & Virology
                Keywords: autophagy machinery, pedv replication, inflammatory responses, apoptosis

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