Functional organization of cytoplasmic inclusion bodies in cells infected by respiratory syncytial virus

Rincheval, Vincent; Lelek, Mickaël; Gault, Elyanne; Bouillier, Camille; Sitterlin, Delphine; Blouquit-Laye, Sabine; Galloux, Marie; Zimmer, Christophe; Éléouët, Jean-François; Rameix‐Welti, Marie‐Anne

doi:10.1038/s41467-017-00655-9

Cited by 160 publications

(261 citation statements)

References 60 publications

(95 reference statements)

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“…In Mononegavirales, transcription and replication takes place in viral factories, e.g. cytoplasmic inclusions where the local concentration of viral proteins is increased thereby facilitating viral replication [53][54][55][56][57][58]. These viral factories, which can be either doublemembrane [59] or membrane-less organelles [60], also likely prevent the activation of host innate immunity by shielding viral proteins and thus impairing their interaction with cellular antiviral proteins.…”

Section: Discussionmentioning

confidence: 99%

Phase transition and amyloid formation by a viral protein as an additional molecular mechanism of virus-induced cell toxicity

Salladini

Debarnot

Delauzun

et al. 2018

Preprint

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Henipaviruses are severe human pathogens responsible for severe encephalitis. Their V protein is a key player in the evasion of the host innate immune response. We have previously reported a biophysical characterization of the Henipavirus V proteins and shown that they interact with DDB1, a cellular protein that is a component of the ubiquitin ligase E3 complex. Here, we serendipitously discovered that the Hendra virus V protein undergoes a liquidhydrogel phase transition. By combining experimental and bioinformatics approaches, we have identified the V region responsible for this phenomenon. This region (referred to as PNT3), which falls within the long intrinsically disordered region of V, was further investigated using a combination of biophysical and structural approaches. ThioflavinT and Congo red binding assays, together with negative-staining electron microscopy studies, show that this region forms amyloid-like, β-enriched structures. Such structures are also formed in mammal cells transfected to express PNT3. Those cells also exhibit a reduced viability in the presence of a stress agent. Interestingly, mammal cells expressing a rationally designed, nonamyloidogenic PNT3 variant (PNT3 3A ), appear to be much less sensitive to the stress agent, thus enabling the establishment of a link between fibril formation and cell toxicity. The present findings therefore pinpoint a so far never reported possible mechanism of virusinduced cell toxicity.

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Section: Discussionmentioning

confidence: 99%

Phase transition and amyloid formation by a viral protein as an additional molecular mechanism of virus-induced cell toxicity

Salladini

Debarnot

Delauzun

et al. 2018

Preprint

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“…After fusion, the helical ribonucleoprotein complex (RNP) is released into the host cell cytoplasm. Transcription and replication occur in the cytoplasm in viral inclusion bodies that serve to concentrate viral products [165][166][167] . The viral RNA-dependent RNA polymerase (RdRp) complex is responsible for transcribing viral mRNA and synthesizing positive-sense anti-genome intermediates needed for replication of new negative-sense genomes for packaging into virions (reviewed elsewhere 168 ).…”

Section: Box 1 | the Life Cycle Of Respiratory Syncytial Virusmentioning

confidence: 99%

Respiratory syncytial virus entry and how to block it

2019

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AlveoliSmall air sacs in the lung that provide rapid gas exchange with blood. BronchiolitisInflammation of the bronchioles that reduces air passage. FormalinAn aqueous solution of formaldehyde.Abstract | Respiratory syncytial virus (RSV) is a leading cause of lower respiratory tract disease in young children and elderly people. Although the virus was isolated in 1955, an effective RSV vaccine has not been developed, and the only licensed intervention is passive immunoprophylaxis of high-risk infants with a humanized monoclonal antibody. During the past 5 years, however, there has been substantial progress in our understanding of the structure and function of the RSV glycoproteins and their interactions with host cell factors that mediate entry. This period has coincided with renewed interest in developing effective interventions, including the isolation of potent monoclonal antibodies and small molecules and the design of novel vaccine candidates. In this Review , we summarize the recent findings that have begun to elucidate RSV entry mechanisms, describe progress on the development of new interventions and conclude with a perspective on gaps in our knowledge that require further investigation.

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“…In our siRNA screen we showed that the nuclear RNA export factor 1 (NXF1) is necessary for the EBOV life cycle, and also for the life cycle of the highly pathogenic New World arenavirus Junín virus. These data suggest a mechanism of action that may be conserved among several cytoplasmically replicating negative-stranded RNA viruses (NSVs) that share commonalties in their replication cycles, such as replication in cytoplasmic inclusion bodies [10,[21][22][23][24]. Importantly, we also could show that NXF1 is important for the life cycle of EBOV in context of infectious virus [19].…”

Section: Introductionmentioning

confidence: 74%

“…A second challenge during the EBOV life cycle is the fact that viral RNA synthesis occurs in inclusion bodies. These are increasingly appreciated for their importance in the genome replication of cytoplasmically replicating NSVs [12,[21][22][23]51], and have for some of these viruses been described to show properties of liquid organelles [24,52]. Similar properties have been observed for EBOV inclusion bodies [10], suggesting that inclusion bodies of NSVs might generally represent liquid organelles.…”

Section: A Model For the Function Of Nxf1 In The Ebov Life Cyclementioning

confidence: 80%

The Ebola Virus Nucleoprotein Recruits the Nuclear RNA Export Factor NXF1 into Inclusion Bodies to Facilitate Viral Protein Expression

Wendt

Brandt

Bodmer

et al. 2020

Cells

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Ebola virus (EBOV) causes severe outbreaks of viral hemorrhagic fever in humans. While virus-host interactions are promising targets for antivirals, there is only limited knowledge regarding the interactions of EBOV with cellular host factors. Recently, we performed a genome-wide siRNA screen that identified the nuclear RNA export factor 1 (NXF1) as an important host factor for the EBOV life cycle. NXF1 is a major component of the nuclear mRNA export pathway that is usurped by many viruses whose life cycles include nuclear stages. However, the role of NXF1 in the life cycle of EBOV, a virus replicating in cytoplasmic inclusion bodies, remains unknown. In order to better understand the role of NXF1 in the EBOV life cycle, we performed a combination of co-immunoprecipitation and double immunofluorescence assays to characterize the interactions of NXF1 with viral proteins and RNAs. Additionally, using siRNA-mediated knockdown of NXF1 together with functional assays, we analyzed the role of NXF1 in individual aspects of the virus life cycle. With this approach we identified the EBOV nucleoprotein (NP) as a viral interaction partner of NXF1. Further studies revealed that NP interacts with the RNA-binding domain of NXF1 and competes with RNA for this interaction. Co-localization studies showed that RNA binding-deficient, but not wildtype NXF1, accumulates in NP-derived inclusion bodies, and knockdown experiments demonstrated that NXF1 is necessary for viral protein expression, but not for viral RNA synthesis. Finally, our results showed that NXF1 interacts with viral mRNAs, but not with viral genomic RNAs. Based on these results we suggest a model whereby NXF1 is recruited into inclusion bodies to promote the export of viral mRNA:NXF1 complexes from these sites. This would represent a novel function for NXF1 in the life cycle of cytoplasmically replicating viruses, and may provide a basis for new therapeutic approaches against EBOV, and possibly other emerging viruses.

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Functional organization of cytoplasmic inclusion bodies in cells infected by respiratory syncytial virus

Cited by 160 publications

References 60 publications

Phase transition and amyloid formation by a viral protein as an additional molecular mechanism of virus-induced cell toxicity

Phase transition and amyloid formation by a viral protein as an additional molecular mechanism of virus-induced cell toxicity

Respiratory syncytial virus entry and how to block it

The Ebola Virus Nucleoprotein Recruits the Nuclear RNA Export Factor NXF1 into Inclusion Bodies to Facilitate Viral Protein Expression

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