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Adenovirus vector-based genetic vaccines have emerged as a powerful strategy against the SARS-CoV-2 health crisis. This success is not unexpected because adenoviruses combine many desirable features of a genetic vaccine. They are highly immunogenic and have a low and well characterized pathogenic profile paired with technological approachability. Ongoing efforts to improve adenovirus-vaccine vectors include the use of rare serotypes and non-human adenoviruses. In this review, we focus on the viral capsid and how the choice of genotypes influences the uptake and subsequent subcellular sorting. We describe how understanding capsid properties, such as stability during the entry process, can change the fate of the entering particles and how this translates into differences in immunity outcomes. We discuss in detail how mutating the membrane lytic capsid protein VI affects species C viruses’ post-entry sorting and briefly discuss if such approaches could have a wider implication in vaccine and/or vector development.
Coralie Daussy; Noémie Pied; Harald Wodrich. Understanding Post Entry Sorting of Adenovirus Capsids; A Chance to Change Vaccine Vector Properties. Viruses 2021, 13, 1221 .
AMA StyleCoralie Daussy, Noémie Pied, Harald Wodrich. Understanding Post Entry Sorting of Adenovirus Capsids; A Chance to Change Vaccine Vector Properties. Viruses. 2021; 13 (7):1221.
Chicago/Turabian StyleCoralie Daussy; Noémie Pied; Harald Wodrich. 2021. "Understanding Post Entry Sorting of Adenovirus Capsids; A Chance to Change Vaccine Vector Properties." Viruses 13, no. 7: 1221.
The beta-coronavirus SARS-CoV-2 is at the origin of a persistent worldwide pandemic. SARS-CoV-2 infections initiate in the bronchi of the upper respiratory tract and are able to disseminate to the lower respiratory tract eventually causing acute severe respiratory syndrome with a high degree of mortality in the elderly. Here we use reconstituted primary bronchial epithelia from adult and children donors to follow the infection dynamic following infection with SARS-CoV-2. We show that in bronchial epithelia derived from adult donors, infections initiate in multi-ciliated cells. Then, infection rapidly spread within 24-48h throughout the whole epithelia. Within 3-4 days, large apical syncytia form between multi-ciliated cells and basal cells, which dissipate into the apical lumen. We show that these syncytia are a significant source of the released infectious dose. In stark contrast to these findings, bronchial epithelia reconstituted from children donors are intrinsically more resistant to virus infection and show active restriction of virus spread. This restriction is paired with accelerated release of IFN compared to adult donors. Taken together our findings reveal apical syncytia formation as an underappreciated source of infectious virus for either local dissemination or release into the environment. Furthermore, we provide direct evidence that children bronchial epithelia are more resistant to infection with SARS-CoV-2 providing experimental support for epidemiological observations that SARS-CoV-2 cases’ fatality is linked to age. Significance Statement Bronchial epithelia are the primary target for SARS-CoV-2 infections. Our work uses reconstituted bronchial epithelia from adults and children. We show that infection of adult epithelia with SARS-CoV-2 is rapid and results in the synchronized release of large clusters of infected cells and syncytia into the apical lumen contributing to the released infectious virus dose. Infection of children derived bronchial epithelia revealed an intrinsic resistance to infection and virus spread, probably as a result of a faster onset of interferon secretion. Thus, our data provide direct evidence for the epidemiological observation that children are less susceptible to SARS-CoV-2.
Guillaume Beucher; Marie-Lise Blondot; Alexis Celle; Noémie Pied; Patricia Recordon-Pinson; Pauline Esteves; Muriel Faure; Mathieu Métifiot; Sabrina Lacomme; Denis Dacheaux; Derrick Robinson; Gernot Längst; Fabien Beaufils; Marie-Edith Lafon; Patrick Berger; Marc Landry; Denis Malvy; Thomas Trian; Marie-Line Andreola; Harald Wodrich. SARS-CoV-2 transmission via apical syncytia release from primary bronchial epithelia and infectivity restriction in children epithelia. 2021, 1 .
AMA StyleGuillaume Beucher, Marie-Lise Blondot, Alexis Celle, Noémie Pied, Patricia Recordon-Pinson, Pauline Esteves, Muriel Faure, Mathieu Métifiot, Sabrina Lacomme, Denis Dacheaux, Derrick Robinson, Gernot Längst, Fabien Beaufils, Marie-Edith Lafon, Patrick Berger, Marc Landry, Denis Malvy, Thomas Trian, Marie-Line Andreola, Harald Wodrich. SARS-CoV-2 transmission via apical syncytia release from primary bronchial epithelia and infectivity restriction in children epithelia. . 2021; ():1.
Chicago/Turabian StyleGuillaume Beucher; Marie-Lise Blondot; Alexis Celle; Noémie Pied; Patricia Recordon-Pinson; Pauline Esteves; Muriel Faure; Mathieu Métifiot; Sabrina Lacomme; Denis Dacheaux; Derrick Robinson; Gernot Längst; Fabien Beaufils; Marie-Edith Lafon; Patrick Berger; Marc Landry; Denis Malvy; Thomas Trian; Marie-Line Andreola; Harald Wodrich. 2021. "SARS-CoV-2 transmission via apical syncytia release from primary bronchial epithelia and infectivity restriction in children epithelia." , no. : 1.
Cells are constantly challenged by pathogens (bacteria, virus, and fungi), and protein aggregates or chemicals, which can provoke membrane damage at the plasma membrane or within the endo-lysosomal compartments. Detection of endo-lysosomal rupture depends on a family of sugar-binding lectins, known as galectins, which sense the abnormal exposure of glycans to the cytoplasm upon membrane damage. Galectins in conjunction with other factors orchestrate specific membrane damage responses such as the recruitment of the endosomal sorting complex required for transport (ESCRT) machinery to either repair damaged membranes or the activation of autophagy to remove membrane remnants. If not controlled, membrane damage causes the release of harmful components including protons, reactive oxygen species, or cathepsins that will elicit inflammation. In this review, we provide an overview of current knowledge on membrane damage and cellular responses. In particular, we focus on the endo-lysosomal damage triggered by non-enveloped viruses (such as adenovirus) and discuss viral strategies to control the cellular membrane damage response. Finally, we debate the link between autophagy and inflammation in this context and discuss the possibility that virus induced autophagy upon entry limits inflammation.
Coralie F. Daussy; Harald Wodrich. “Repair Me if You Can”: Membrane Damage, Response, and Control from the Viral Perspective. Cells 2020, 9, 2042 .
AMA StyleCoralie F. Daussy, Harald Wodrich. “Repair Me if You Can”: Membrane Damage, Response, and Control from the Viral Perspective. Cells. 2020; 9 (9):2042.
Chicago/Turabian StyleCoralie F. Daussy; Harald Wodrich. 2020. "“Repair Me if You Can”: Membrane Damage, Response, and Control from the Viral Perspective." Cells 9, no. 9: 2042.
Recently an increasing number of new adenovirus types associated with type-dependent pathogenicity have been identified. However, identification of these clinical isolates represents the very first step to characterize novel pathogens. For deeper analyses, these adenoviruses need to be further characterized in basic virology experiments or they could be applied in translational research. To achieve this goal, it is essential to get genetic access and to enable genetic modification of these novel adenovirus genomes (deletion, insertion, and mutation). Here we demonstrate a high-throughput approach to get genetic access to new adenoviruses via homologous recombination. We first defined the cloning conditions regarding homology arm-length and input adenoviral genome amounts. Then we cloned four naturally occurring adenoviruses (Ad70, Ad73, Ad74, and Ad75) into easy-to-manipulate plasmids and genetically modified them by reporter gene insertion. Three recombinant adenoviruses (Ad70, Ad73, and Ad74) containing a reporter cassette were successfully reconstituted. These novel reporter-labeled adenoviruses were further characterized using the inserted luciferase reporter with respect to receptor usage, presence of anti-adenovirus antibodies, and tropism in vitro. The identified receptor usage, the relatively low prevalence of anti-adenovirus antibodies, and the various cancer cell line transduction pattern are important features of these new pathogens providing essential information for their therapeutic application.
Wenli Zhang; Kemal Mese; Sebastian Schellhorn; Nora Bahlmann; Nicolas Mach; Oskar Bunz; Akshay Dhingra; Elias Hage; Marie-Edith Lafon; Harald Wodrich; Albert Heim; Anja Ehrhardt. High-Throughput Cloning and Characterization of Emerging Adenovirus Types 70, 73, 74, and 75. International Journal of Molecular Sciences 2020, 21, 6370 .
AMA StyleWenli Zhang, Kemal Mese, Sebastian Schellhorn, Nora Bahlmann, Nicolas Mach, Oskar Bunz, Akshay Dhingra, Elias Hage, Marie-Edith Lafon, Harald Wodrich, Albert Heim, Anja Ehrhardt. High-Throughput Cloning and Characterization of Emerging Adenovirus Types 70, 73, 74, and 75. International Journal of Molecular Sciences. 2020; 21 (17):6370.
Chicago/Turabian StyleWenli Zhang; Kemal Mese; Sebastian Schellhorn; Nora Bahlmann; Nicolas Mach; Oskar Bunz; Akshay Dhingra; Elias Hage; Marie-Edith Lafon; Harald Wodrich; Albert Heim; Anja Ehrhardt. 2020. "High-Throughput Cloning and Characterization of Emerging Adenovirus Types 70, 73, 74, and 75." International Journal of Molecular Sciences 21, no. 17: 6370.
Inside the capsid adenovirus DNA is associated with the structural protein pVII. However, the viral DNA organisation, pVII positioning and the dynamic changes of viral packaging upon infection, to form a transcriptional active genome, are not known. We combined MNase-Seq and single genome imaging during early infection to provide the structure and time resolved dynamics of viral chromatin changes, correlated with gene transcription. pVII complexes form nucleosome-like arrays, being precisely positioned on DNA, creating a defined and unique adenoviral nucleoprotein-architecture. The structure renders the viral genome transcription competent with lower pVII densities at early gene loci, correlating with viral chromatin de-condensation upon infection. Nucleosomes specifically replace pVII at transcription start sites of early genes, preceding transcriptional activation. Our study suggests an underlying regulatory pVII nucleoprotein-architecture, required for the dynamic changes during early infection, including transcription related nucleosome assembly. We suggest that our study provides a basis for the development of recombinant adenoviral vectors exhibiting sustained expression in gene therapy.
Uwe Schwartz; Tetsuro Komatsu; Claudia Huber; Floriane Lagadec; Elisabeth Silberhorn; Edouard Bertrand; Harald Wodrich; Gernot Laengst. A pVII positioning code determines the dynamics of Adenoviral nucleoprotein structure and primes it for early gene activation. 2020, 1 .
AMA StyleUwe Schwartz, Tetsuro Komatsu, Claudia Huber, Floriane Lagadec, Elisabeth Silberhorn, Edouard Bertrand, Harald Wodrich, Gernot Laengst. A pVII positioning code determines the dynamics of Adenoviral nucleoprotein structure and primes it for early gene activation. . 2020; ():1.
Chicago/Turabian StyleUwe Schwartz; Tetsuro Komatsu; Claudia Huber; Floriane Lagadec; Elisabeth Silberhorn; Edouard Bertrand; Harald Wodrich; Gernot Laengst. 2020. "A pVII positioning code determines the dynamics of Adenoviral nucleoprotein structure and primes it for early gene activation." , no. : 1.
Nuclear import of viral genomes is an essential step to initiate productive infection for several nuclear replicating DNA viruses. On the other hand, DNA is not a physiological nuclear import substrate; consequently, viruses have to exploit existing physiological transport routes. Here, we show that adenoviruses use the nucleoporin Nup358 to increase the efficiency of adenoviral genome import. In its absence, genome import efficiency is reduced and the transport receptor transportin 1 becomes rate limiting. We show that the N-terminal half of Nup358 is sufficient to drive genome import and identify a transportin 1 binding region. In our model, adenovirus genome import exploits an existing protein import pathway and Nup358 serves as an assembly platform for transport complexes.
Irene Carlon-Andres; Floriane Lagadec; Noémie Pied; Fabienne Rayne; Marie-Edith Lafon; Ralph H. Kehlenbach; Harald Wodrich. Nup358 and Transportin 1 Cooperate in Adenoviral Genome Import. Journal of Virology 2020, 94, 1 .
AMA StyleIrene Carlon-Andres, Floriane Lagadec, Noémie Pied, Fabienne Rayne, Marie-Edith Lafon, Ralph H. Kehlenbach, Harald Wodrich. Nup358 and Transportin 1 Cooperate in Adenoviral Genome Import. Journal of Virology. 2020; 94 (10):1.
Chicago/Turabian StyleIrene Carlon-Andres; Floriane Lagadec; Noémie Pied; Fabienne Rayne; Marie-Edith Lafon; Ralph H. Kehlenbach; Harald Wodrich. 2020. "Nup358 and Transportin 1 Cooperate in Adenoviral Genome Import." Journal of Virology 94, no. 10: 1.
Incoming adenoviruses seize control of cytosolic transport mechanisms to relocate their genome from the cell periphery to specialized sites in the nucleoplasm. The nucleus is the site for viral gene expression, genome replication and the production of progeny for the next round of infection. By taking control of the cell, adenoviruses also suppress cell autonomous immunity responses. To succeed in their production cycle, adenoviruses rely on well‐coordinated steps, facilitated by interactions between viral proteins and cellular factors. Interactions between virus and host can impose remarkable morphological changes in the infected cell. Imaging adenoviruses has tremendously influenced how we delineate individual steps in the viral life cycle, because it allowed the development of specific optical markers to label these morphological changes in space and time. As technology advances, innovative imaging techniques and novel tools for specimen labeling keeps uncovering previously unseen facets of adenovirus biology emphasizing why imaging adenoviruses is as attractive today as it was in the past. This review will summarize past achievements and present developments in adenovirus imaging centered on fluorescence microscopy approaches.
Noémie Pied; Harald Wodrich. Imaging the adenovirus infection cycle. FEBS Letters 2019, 593, 3419 -3448.
AMA StyleNoémie Pied, Harald Wodrich. Imaging the adenovirus infection cycle. FEBS Letters. 2019; 593 (24):3419-3448.
Chicago/Turabian StyleNoémie Pied; Harald Wodrich. 2019. "Imaging the adenovirus infection cycle." FEBS Letters 593, no. 24: 3419-3448.
The adenovirus (Ad) genome is believed to be packaged into the virion by forming a chromatin‐like structure. The replicated viral genome is likely to be condensed through binding with viral core proteins before encapsidation. Replicated viral genomes accumulate in the central region of the nucleus, which we termed virus‐induced postreplication (ViPR) body. However, the molecular mechanism by which the nuclear structure is reorganized and its functional significance in virus production are currently not understood. In this study, we found that viral packaging protein IVa2, but not capsid proteins, accumulated in the ViPR body. In addition, nucleolar chromatin regulatory proteins, nucleophosmin 1 (NPM1), upstream binding factor, and nucleolin accumulated in the ViPR body in late‐stage Ad infection. NPM1 depletion increased the nuclease‐resistant viral genome and delayed the ViPR body formation. These results suggested that structural changes in the infected cell nucleus depend on the formation of viral chromatin by host chromatin regulatory proteins. Because NPM1 depletion decreases production of the infectious virion, we propose that host factor‐mediated viral chromatin remodeling and concomitant ViPR body formation are prerequisites for efficient encapsidation of Ad chromatin.
Michelle Jane Genoveso; Miharu Hisaoka; Tetsuro Komatsu; Harald Wodrich; Kyosuke Nagata; Mitsuru Okuwaki. Formation of adenovirus DNA replication compartments and viral DNA accumulation sites by host chromatin regulatory proteins including NPM1. The FEBS Journal 2019, 287, 205 -217.
AMA StyleMichelle Jane Genoveso, Miharu Hisaoka, Tetsuro Komatsu, Harald Wodrich, Kyosuke Nagata, Mitsuru Okuwaki. Formation of adenovirus DNA replication compartments and viral DNA accumulation sites by host chromatin regulatory proteins including NPM1. The FEBS Journal. 2019; 287 (1):205-217.
Chicago/Turabian StyleMichelle Jane Genoveso; Miharu Hisaoka; Tetsuro Komatsu; Harald Wodrich; Kyosuke Nagata; Mitsuru Okuwaki. 2019. "Formation of adenovirus DNA replication compartments and viral DNA accumulation sites by host chromatin regulatory proteins including NPM1." The FEBS Journal 287, no. 1: 205-217.
Chronic infection with the human Hepatitis B virus (HBV) is a major global health problem. Hepatitis D virus (HDV) is a satellite of HBV that uses HBV envelope proteins for cell egress and entry. Using infection systems encoding the HBV/HDV receptor human sodium taurocholate co-transporting polypeptide (NTCP), we screened 1181 FDA-approved drugs applying markers for interference for HBV and HDV infection. As one primary hit we identified Acitretin, a retinoid, as an inhibitor of HBV replication and HDV release. Based on this, other retinoic acid receptor (RAR) agonists with different specificities were found to interfere with HBV replication, verifying that the retinoic acid receptor pathway regulates replication. Of the eight agonists investigated, RARα-specific agonist Am80 (tamibarotene) was most active. Am80 reduced secretion of HBeAg and HBsAg with IC50s < 10 nM in differentiated HepaRG-NTCP cells. Similar effects were observed in primary human hepatocytes. In HepG2-NTCP cells, profound Am80-mediated inhibition required prolonged treatment of up to 35 days. Am80 treatment of cells with an established HBV cccDNA pool resulted in a reduction of secreted HBsAg and HBeAg, which correlated with reduced intracellular viral RNA levels, but not cccDNA copy numbers. The effect lasted for >12 days after removal of the drug. HBV genotypes B, D, and E were equally inhibited. By contrast, Am80 did not affect HBV replication in transfected cells or HepG2.2.15 cells, which carry an integrated HBV genome. In conclusion, our results indicate a persistent inhibition of HBV transcription by Am80, which might be used for drug repositioning.
Shirin Nkongolo; Lea Nußbaum; Florian A. Lempp; Harald Wodrich; Stephan Urban; Yi Ni. The retinoic acid receptor (RAR) α-specific agonist Am80 (tamibarotene) and other RAR agonists potently inhibit hepatitis B virus transcription from cccDNA. Antiviral Research 2019, 168, 146 -155.
AMA StyleShirin Nkongolo, Lea Nußbaum, Florian A. Lempp, Harald Wodrich, Stephan Urban, Yi Ni. The retinoic acid receptor (RAR) α-specific agonist Am80 (tamibarotene) and other RAR agonists potently inhibit hepatitis B virus transcription from cccDNA. Antiviral Research. 2019; 168 ():146-155.
Chicago/Turabian StyleShirin Nkongolo; Lea Nußbaum; Florian A. Lempp; Harald Wodrich; Stephan Urban; Yi Ni. 2019. "The retinoic acid receptor (RAR) α-specific agonist Am80 (tamibarotene) and other RAR agonists potently inhibit hepatitis B virus transcription from cccDNA." Antiviral Research 168, no. : 146-155.
Viruses must deliver their genomes to host cells to ensure replication and propagation. Characterizing the fate of viral genomes is crucial to understand the viral life cycle and the fate of virus-derived vector tools. Here, we integrated the ANCHOR3 system, an in vivo DNA-tagging technology, into the adenoviral genome for real-time genome detection. ANCHOR3 tagging permitted the in vivo visualization of incoming genomes at the onset of infection and of replicated genomes at late phases of infection. Using this system, we show viral genome attachment to condensed host chromosomes during mitosis, identifying this mechanism as a mode of cell-to-cell transfer. We characterize the spatiotemporal organization of adenovirus replication and identify two kinetically distinct phases of viral genome replication. The ANCHOR3 system is the first technique that allows the continuous visualization of adenoviral genomes during the entire virus life cycle, opening the way for further in-depth study.
Tetsuro Komatsu; Charlotte Quentin-Froignant; Irene Carlon-Andres; Floriane Lagadec; Fabienne Rayne; Jessica Ragues; Ralph H. Kehlenbach; Wenli Zhang; Anja Ehrhardt; Kerstin Bystricky; Renaud Morin; Jean-Michel Lagarde; Franck Gallardo; Harald Wodrich. In Vivo Labelling of Adenovirus DNA Identifies Chromatin Anchoring and Biphasic Genome Replication. Journal of Virology 2018, 92, 1 .
AMA StyleTetsuro Komatsu, Charlotte Quentin-Froignant, Irene Carlon-Andres, Floriane Lagadec, Fabienne Rayne, Jessica Ragues, Ralph H. Kehlenbach, Wenli Zhang, Anja Ehrhardt, Kerstin Bystricky, Renaud Morin, Jean-Michel Lagarde, Franck Gallardo, Harald Wodrich. In Vivo Labelling of Adenovirus DNA Identifies Chromatin Anchoring and Biphasic Genome Replication. Journal of Virology. 2018; 92 (18):1.
Chicago/Turabian StyleTetsuro Komatsu; Charlotte Quentin-Froignant; Irene Carlon-Andres; Floriane Lagadec; Fabienne Rayne; Jessica Ragues; Ralph H. Kehlenbach; Wenli Zhang; Anja Ehrhardt; Kerstin Bystricky; Renaud Morin; Jean-Michel Lagarde; Franck Gallardo; Harald Wodrich. 2018. "In Vivo Labelling of Adenovirus DNA Identifies Chromatin Anchoring and Biphasic Genome Replication." Journal of Virology 92, no. 18: 1.
Adenoviruses are characterized by a large variability, reflected by their classification in species A to G. Certain species, eg A and C, could be associated with increased clinical severity, both in immunocompetent and immunocompromised hosts suggesting that in some instances species identification provides clinically relevant information. Here we designed a novel "pVI rapid typing method" to obtain quick, simple and cost effective species assignment for Adenoviruses, thanks to combined fusion temperature (Tm) and amplicon size analysis. Rapid typing results were compared to Sanger sequencing in the hexon gene for 140 Adenovirus-positive clinical samples included in the Typadeno study. Species A and C could be identified with a 100% positive predictive value, thus confirming the value of this simple typing method.
Fabienne Rayne; Linda Wittkop; Clément Bader; Somar Kassab; Camille Tumiotto; Sylvie Berciaud; Harald Wodrich; Marie-Edith Lafon. Rapid Adenovirus typing method for species identification. Journal of Virological Methods 2017, 249, 156 -160.
AMA StyleFabienne Rayne, Linda Wittkop, Clément Bader, Somar Kassab, Camille Tumiotto, Sylvie Berciaud, Harald Wodrich, Marie-Edith Lafon. Rapid Adenovirus typing method for species identification. Journal of Virological Methods. 2017; 249 ():156-160.
Chicago/Turabian StyleFabienne Rayne; Linda Wittkop; Clément Bader; Somar Kassab; Camille Tumiotto; Sylvie Berciaud; Harald Wodrich; Marie-Edith Lafon. 2017. "Rapid Adenovirus typing method for species identification." Journal of Virological Methods 249, no. : 156-160.
Autophagy is an essential metabolic program that is also used for clearing intracellular pathogens. This mechanism, also termed selective autophagy, is well characterized for invasive bacteria but remains poorly documented for viral infections. Here we highlight our recent work showing that endosomolytic adenoviruses trigger autophagy when entering cells. Our study revealed how adenoviruses exploit a capsid-associated small PPxY peptide motif to manipulate the autophagic machinery to prevent autophagic degradation and to promote endosomal escape and nuclear trafficking.
Charlotte Montespan; Christopher M. Wiethoff; Harald Wodrich. A Small Viral PPxY Peptide Motif To Control Antiviral Autophagy. Journal of Virology 2017, 91, 1 .
AMA StyleCharlotte Montespan, Christopher M. Wiethoff, Harald Wodrich. A Small Viral PPxY Peptide Motif To Control Antiviral Autophagy. Journal of Virology. 2017; 91 (18):1.
Chicago/Turabian StyleCharlotte Montespan; Christopher M. Wiethoff; Harald Wodrich. 2017. "A Small Viral PPxY Peptide Motif To Control Antiviral Autophagy." Journal of Virology 91, no. 18: 1.
Cells employ active measures to restrict infection by pathogens, even prior to responses from the innate and humoral immune defenses. In this context selective autophagy is activated upon pathogen induced membrane rupture to sequester and deliver membrane fragments and their pathogen contents for lysosomal degradation. Adenoviruses, which breach the endosome upon entry, escape this fate by penetrating into the cytosol prior to autophagosome sequestration of the ruptured endosome. We show that virus induced membrane damage is recognized through Galectin-8 and sequesters the autophagy receptors NDP52 and p62. We further show that a conserved PPxY motif in the viral membrane lytic protein VI is critical for efficient viral evasion of autophagic sequestration after endosomal lysis. Comparing the wildtype with a PPxY-mutant virus we show that depletion of Galectin-8 or suppression of autophagy in ATG5-/- MEFs rescues infectivity of the PPxY-mutant virus while depletion of the autophagy receptors NDP52, p62 has only minor effects. Furthermore we show that wildtype viruses exploit the autophagic machinery for efficient nuclear genome delivery and control autophagosome formation via the cellular ubiquitin ligase Nedd4.2 resulting in reduced antigenic presentation. Our data thus demonstrate that a short PPxY-peptide motif in the adenoviral capsid permits multi-layered viral control of autophagic processes during entry. Cells have developed surveillance systems to detect invading pathogens, e.g. when they damage the membrane upon cell entry. Cells respond to membrane damage by activating selective autophagy to target pathogens for lysosomal degradation and pathogen removal. In this manuscript we show that endosome penetrating adenoviruses also activate an autophagic response upon entry and that this involves Galectin-8 mediated membrane damage recognition similar to pathogenic bacteria. In contrast adenoviruses avoid degradation by escaping into the cytosol. We show that they possess a small virion encoded PPxY-peptide motif, which they use to divert the autophagic response of the cell. This motif sequesters the cellular ubiquitin ligase Nedd4.2 resulting in limited autophagosome formation, which prevents viral degradation and antigenic presentation and ensures efficient endosomal escape and nuclear transport.
Charlotte Montespan; Shauna A. Marvin; Sisley Austin; Andrew M. Burrage; Benoit Roger; Fabienne Rayne; Muriel Faure; Edward M. Campell; Carola Schneider; Rudolph Reimer; Kay Grünewald; Christopher M. Wiethoff; Harald Wodrich. Multi-layered control of Galectin-8 mediated autophagy during adenovirus cell entry through a conserved PPxY motif in the viral capsid. PLOS Pathogens 2017, 13, e1006217 .
AMA StyleCharlotte Montespan, Shauna A. Marvin, Sisley Austin, Andrew M. Burrage, Benoit Roger, Fabienne Rayne, Muriel Faure, Edward M. Campell, Carola Schneider, Rudolph Reimer, Kay Grünewald, Christopher M. Wiethoff, Harald Wodrich. Multi-layered control of Galectin-8 mediated autophagy during adenovirus cell entry through a conserved PPxY motif in the viral capsid. PLOS Pathogens. 2017; 13 (2):e1006217.
Chicago/Turabian StyleCharlotte Montespan; Shauna A. Marvin; Sisley Austin; Andrew M. Burrage; Benoit Roger; Fabienne Rayne; Muriel Faure; Edward M. Campell; Carola Schneider; Rudolph Reimer; Kay Grünewald; Christopher M. Wiethoff; Harald Wodrich. 2017. "Multi-layered control of Galectin-8 mediated autophagy during adenovirus cell entry through a conserved PPxY motif in the viral capsid." PLOS Pathogens 13, no. 2: e1006217.
Unr is a cytoplasmic RNA-binding protein with roles in the regulation of mRNA stability and translation. In this study, we have identified a novel function for Unr, which acts as a positive regulator of placental development. Unr expression studies in the developing placenta revealed the presence of Unr-rich foci apparently localized in the nuclei of trophoblast giant cells (TGCs). We determined that these foci are actually cross sections of a network of double-wall nuclear membranes invaginations containing a cytoplasmic core, related to the nucleoplasmic reticulum (NR), and accordingly named Unr-NRs. Unr-NRs constitute a novel type of NR, because they contain high levels of poly (A) RNA and translation factors and are sites of active translation. In murine tissues, Unr-NRs are only found in two polyploid cell types, TGCs and hepatocytes. In vitro, their formation is linked to stress and polyploidy, since in three cancer cell lines, cytotoxic drugs known to promote polyploidization induce their formation. Finally, we show that Unr is required, in vivo, for Unr-NRs formation, because these structures are lacking in Unr-null TGCs.
Frédéric Saltel; Alban Giese; Lamia Azzi-Martin; Habiba Elatmani; Pierre Costet; Zakaria Ezzoukhry; Nathalie Dugot-Senant; Lucile Miquerol; Oréda Boussadia; Harald Wodrich; Pierre Dubus; Hélène Jacquemin-Sablon. Unr defines a novel class of nucleoplasmic reticulum, involved in mRNA translation. Journal of Cell Science 2017, 130, 1796 -1808.
AMA StyleFrédéric Saltel, Alban Giese, Lamia Azzi-Martin, Habiba Elatmani, Pierre Costet, Zakaria Ezzoukhry, Nathalie Dugot-Senant, Lucile Miquerol, Oréda Boussadia, Harald Wodrich, Pierre Dubus, Hélène Jacquemin-Sablon. Unr defines a novel class of nucleoplasmic reticulum, involved in mRNA translation. Journal of Cell Science. 2017; 130 (10):1796-1808.
Chicago/Turabian StyleFrédéric Saltel; Alban Giese; Lamia Azzi-Martin; Habiba Elatmani; Pierre Costet; Zakaria Ezzoukhry; Nathalie Dugot-Senant; Lucile Miquerol; Oréda Boussadia; Harald Wodrich; Pierre Dubus; Hélène Jacquemin-Sablon. 2017. "Unr defines a novel class of nucleoplasmic reticulum, involved in mRNA translation." Journal of Cell Science 130, no. 10: 1796-1808.
In recent years, it has been suggested that host cells exert intrinsic mechanisms to control nuclear replicating DNA viruses. This cellular response involves nuclear antiviral factors targeting incoming viral genomes. Herpes simplex virus-1 (HSV-1) is the best-studied model in this context, and it was shown that upon nuclear entry HSV-1 genomes are immediately targeted by components of promyelocytic leukemia nuclear bodies (PML-NBs) and the nuclear DNA sensor IFI16 (interferon gamma inducible protein 16). Based on HSV-1 studies, together with limited examples in other viral systems, these phenomena are widely believed to be a common cellular response to incoming viral genomes, although formal evidence for each virus is lacking. Indeed, recent studies suggest that the case may be different for adenovirus infection. Here we summarize the existing experimental evidence for the roles of nuclear antiviral factors against incoming viral genomes to better understand cellular responses on a virus-by-virus basis. We emphasize that cells seem to respond differently to different incoming viral genomes and discuss possible arguments for and against a unifying cellular mechanism targeting the incoming genomes of different virus families.
Tetsuro Komatsu; Kyosuke Nagata; Harald Wodrich. The Role of Nuclear Antiviral Factors against Invading DNA Viruses: The Immediate Fate of Incoming Viral Genomes. Viruses 2016, 8, 290 .
AMA StyleTetsuro Komatsu, Kyosuke Nagata, Harald Wodrich. The Role of Nuclear Antiviral Factors against Invading DNA Viruses: The Immediate Fate of Incoming Viral Genomes. Viruses. 2016; 8 (10):290.
Chicago/Turabian StyleTetsuro Komatsu; Kyosuke Nagata; Harald Wodrich. 2016. "The Role of Nuclear Antiviral Factors against Invading DNA Viruses: The Immediate Fate of Incoming Viral Genomes." Viruses 8, no. 10: 290.
AlphaScreen® is a technology particularly suitable for bi-molecular inhibitor screening assays, e.g. using protein–protein interactions with purified recombinant proteins. Each binding partner of the bi-molecular interaction is coupled either to donor or to acceptor beads. The technology is based on the quantifiable transfer of oxygen singlets from donor to acceptor microbeads brought together by a specific interaction between the partners. We identified the conserved interaction between WW domains of cellular ubiquitin ligases of the Nedd4 family and a short peptide motif (PPxY) present in several structural and non-structural viral proteins as a potential drug target. Using an AlphaScreen assay recapitulating the interaction between Nedd4.2 and the PPxY motif of the adenoviral capsid protein VI, we screened a library of small molecules and identified specific inhibitors of this interaction.
Sisley Austin; Saïd Taouji; Eric Chevet; Harald Wodrich; Fabienne Rayne. Using AlphaScreen® to Identify Small-Molecule Inhibitors Targeting a Conserved Host–Pathogen Interaction. Methods in Molecular Biology 2016, 1449, 453 -467.
AMA StyleSisley Austin, Saïd Taouji, Eric Chevet, Harald Wodrich, Fabienne Rayne. Using AlphaScreen® to Identify Small-Molecule Inhibitors Targeting a Conserved Host–Pathogen Interaction. Methods in Molecular Biology. 2016; 1449 ():453-467.
Chicago/Turabian StyleSisley Austin; Saïd Taouji; Eric Chevet; Harald Wodrich; Fabienne Rayne. 2016. "Using AlphaScreen® to Identify Small-Molecule Inhibitors Targeting a Conserved Host–Pathogen Interaction." Methods in Molecular Biology 1449, no. : 453-467.
The CXCL4 paralog CXCL4L1 is a less studied chemokine that has been suggested to exert an antiangiogenic function. However, CXCL4L1 is also expressed in patient tumors, tumor cell lines, and murine xenografts, prompting a more detailed analysis of its role in cancer pathogenesis. We used genetic and antibody-based approaches to attenuate CXCL4L1 in models of pancreatic ductal adenocarcinoma (PDAC). Mechanisms of expression were assessed in cell coculture experiments, murine, and avian xenotransplants, including through an evaluation of CpG methylation and mutation of critical CpG residues. CXCL4L1 gene expression was increased greatly in primary and metastatic PDAC. We found that myofibroblasts triggered cues in the tumor microenvironment, which led to induction of CXCL4L1 in tumor cells. CXCL4L1 expression was also controlled by epigenetic modifications at critical CpG islands, which were mapped. CXCL4L1 inhibited angiogenesis but also affected tumor development more directly, depending on the tumor cell type. In vivo administration of an mAb against CXCL4L1 demonstrated a blockade in the growth of tumors positive for CXCR3, a critical receptor for CXCL4 ligands. Our findings define a protumorigenic role in PDAC development for endogenous CXCL4L1, which is independent of its antiangiogenic function. Cancer Res; 76(22); 6507–19. ©2016 AACR.
Cathy Quemener; Jessica Baud; Kevin Boyé; Alexandre Dubrac; Clotilde Billottet; Fabienne Soulet; Florence Darlot; Laurent Dumartin; Marie Sire; Renaud Grepin; Thomas Daubon; Fabienne Rayne; Harald Wodrich; Anne Couvelard; Raphael Pineau; Martin Schilling; Vincent Castronovo; Shih-Che Sue; Kim Clarke; Abderrahim Lomri; Abdel-Majid Khatib; Martin Hagedorn; Hervé Prats; Andreas Bikfalvi. Dual Roles for CXCL4 Chemokines and CXCR3 in Angiogenesis and Invasion of Pancreatic Cancer. Cancer Research 2016, 76, 6507 -6519.
AMA StyleCathy Quemener, Jessica Baud, Kevin Boyé, Alexandre Dubrac, Clotilde Billottet, Fabienne Soulet, Florence Darlot, Laurent Dumartin, Marie Sire, Renaud Grepin, Thomas Daubon, Fabienne Rayne, Harald Wodrich, Anne Couvelard, Raphael Pineau, Martin Schilling, Vincent Castronovo, Shih-Che Sue, Kim Clarke, Abderrahim Lomri, Abdel-Majid Khatib, Martin Hagedorn, Hervé Prats, Andreas Bikfalvi. Dual Roles for CXCL4 Chemokines and CXCR3 in Angiogenesis and Invasion of Pancreatic Cancer. Cancer Research. 2016; 76 (22):6507-6519.
Chicago/Turabian StyleCathy Quemener; Jessica Baud; Kevin Boyé; Alexandre Dubrac; Clotilde Billottet; Fabienne Soulet; Florence Darlot; Laurent Dumartin; Marie Sire; Renaud Grepin; Thomas Daubon; Fabienne Rayne; Harald Wodrich; Anne Couvelard; Raphael Pineau; Martin Schilling; Vincent Castronovo; Shih-Che Sue; Kim Clarke; Abderrahim Lomri; Abdel-Majid Khatib; Martin Hagedorn; Hervé Prats; Andreas Bikfalvi. 2016. "Dual Roles for CXCL4 Chemokines and CXCR3 in Angiogenesis and Invasion of Pancreatic Cancer." Cancer Research 76, no. 22: 6507-6519.
Fabienne Rayne; Linda Wittkop; Clément Bader; Somar Kassab; Camille Tumiotto; Sylvie Berciaud; Harald Wodrich; Marie-Edith Lafon; Typadeno study members. Rapid adenovirus typing method for species identification. Journal of Clinical Virology 2016, 82, S104 .
AMA StyleFabienne Rayne, Linda Wittkop, Clément Bader, Somar Kassab, Camille Tumiotto, Sylvie Berciaud, Harald Wodrich, Marie-Edith Lafon, Typadeno study members. Rapid adenovirus typing method for species identification. Journal of Clinical Virology. 2016; 82 ():S104.
Chicago/Turabian StyleFabienne Rayne; Linda Wittkop; Clément Bader; Somar Kassab; Camille Tumiotto; Sylvie Berciaud; Harald Wodrich; Marie-Edith Lafon; Typadeno study members. 2016. "Rapid adenovirus typing method for species identification." Journal of Clinical Virology 82, no. : S104.
In adenoviral virions, the genome is organized into a chromatin‐like structure by viral basic core proteins. Consequently viral DNAs must be replicated, chromatinized and packed into progeny virions in infected cells. Although viral DNA replication centers can be visualized by virtue of viral and cellular factors, the spatiotemporal regulation of viral genomes during subsequent steps remains to be elucidated. In this study, we used imaging analyses to examine the fate of adenoviral genomes and to track newly replicated viral DNA as well as replication‐related factors. We show de novo formation of a subnuclear domain, which we termed Virus‐induced Post‐Replication (ViPR) body, that emerges concomitantly with or immediately after disintegration of initial replication centers. Using a nucleoside analogue, we show that viral genomes continue being synthesized in morphologically distinct replication compartments at the periphery of ViPR bodies and are then transported inward. In addition, we identified a nucleolar protein Mybbp1a as a molecular marker for ViPR bodies, which specifically associated with viral core protein VII. In conclusion, our work demonstrates the formation of previously uncharacterized viral DNA replication compartments specific for late phases of infection that produce progeny viral genomes accumulating in ViPR bodies.
Tetsuro Komatsu; Derrick R. Robinson; Miharu Hisaoka; Shuhei Ueshima; Mitsuru Okuwaki; Kyosuke Nagata; Harald Wodrich. Tracking adenovirus genomes identifies morphologically distinct late DNA replication compartments. Traffic 2016, 17, 1168 -1180.
AMA StyleTetsuro Komatsu, Derrick R. Robinson, Miharu Hisaoka, Shuhei Ueshima, Mitsuru Okuwaki, Kyosuke Nagata, Harald Wodrich. Tracking adenovirus genomes identifies morphologically distinct late DNA replication compartments. Traffic. 2016; 17 (11):1168-1180.
Chicago/Turabian StyleTetsuro Komatsu; Derrick R. Robinson; Miharu Hisaoka; Shuhei Ueshima; Mitsuru Okuwaki; Kyosuke Nagata; Harald Wodrich. 2016. "Tracking adenovirus genomes identifies morphologically distinct late DNA replication compartments." Traffic 17, no. 11: 1168-1180.
Recent studies involving several viral systems have highlighted the importance of cellular intrinsic defense mechanisms through nuclear antiviral proteins that restrict viral propagation. These factors include among others components of PML nuclear bodies, the nuclear DNA sensor IFI16, and a potential restriction factor PHF13/SPOC1. For several nuclear replicating DNA viruses, it was shown that these factors sense and target viral genomes immediately upon nuclear import. In contrast to the anticipated view, we recently found that incoming adenoviral genomes are not targeted by PML nuclear bodies. Here we further explored cellular responses against adenoviral infection by focusing on specific conditions as well as additional nuclear antiviral factors. In line with our previous findings, we show that neither interferon treatment nor the use of specific isoforms of PML nuclear body components results in co-localization between incoming adenoviral genomes and the subnuclear domains. Furthermore, our imaging analyses indicated that neither IFI16 nor PHF13/SPOC1 are likely to target incoming adenoviral genomes. Thus our findings suggest that incoming adenoviral genomes may be able to escape from a large repertoire of nuclear antiviral mechanisms, providing a rationale for the efficient initiation of lytic replication cycle.
Tetsuro Komatsu; Hans Will; Kyosuke Nagata; Harald Wodrich. Imaging analysis of nuclear antiviral factors through direct detection of incoming adenovirus genome complexes. Biochemical and Biophysical Research Communications 2016, 473, 200 -205.
AMA StyleTetsuro Komatsu, Hans Will, Kyosuke Nagata, Harald Wodrich. Imaging analysis of nuclear antiviral factors through direct detection of incoming adenovirus genome complexes. Biochemical and Biophysical Research Communications. 2016; 473 (1):200-205.
Chicago/Turabian StyleTetsuro Komatsu; Hans Will; Kyosuke Nagata; Harald Wodrich. 2016. "Imaging analysis of nuclear antiviral factors through direct detection of incoming adenovirus genome complexes." Biochemical and Biophysical Research Communications 473, no. 1: 200-205.