This page has only limited features, please log in for full access.
In the germline of animals, PIWI interacting (pi)RNAs protect the genome against the detrimental effects of transposon mobilization. In Drosophila, piRNA-mediated cleavage of transposon RNA triggers the production of responder piRNAs via ping-pong amplification. Responder piRNA 3′ end formation by the nuclease Zucchini is coupled to the production of downstream trailer piRNAs, expanding the repertoire of transposon piRNA sequences. In Aedes aegypti mosquitoes, piRNAs are generated from viral RNA, yet, it is unknown how viral piRNA 3′ ends are formed and whether viral RNA cleavage gives rise to trailer piRNA production. Here we report that in Ae. aegypti, virus- and transposon-derived piRNAs have sharp 3′ ends, and are biased for downstream uridine residues, features reminiscent of Zucchini cleavage of precursor piRNAs in Drosophila. We designed a reporter system to study viral piRNA 3′ end formation and found that targeting viral RNA by abundant endogenous piRNAs triggers the production of responder and trailer piRNAs. Using this reporter, we identified the Ae. aegypti orthologs of Zucchini and Nibbler, two nucleases involved in piRNA 3′ end formation. Our results furthermore suggest that autonomous piRNA production from viral RNA can be triggered and expanded by an initial cleavage event guided by genome-encoded piRNAs.
Joep Joosten; Gijs J Overheul; Ronald P Van Rij; Pascal Miesen. Endogenous piRNA-guided slicing triggers responder and trailer piRNA production from viral RNA in Aedes aegypti mosquitoes. Nucleic Acids Research 2021, 1 .
AMA StyleJoep Joosten, Gijs J Overheul, Ronald P Van Rij, Pascal Miesen. Endogenous piRNA-guided slicing triggers responder and trailer piRNA production from viral RNA in Aedes aegypti mosquitoes. Nucleic Acids Research. 2021; ():1.
Chicago/Turabian StyleJoep Joosten; Gijs J Overheul; Ronald P Van Rij; Pascal Miesen. 2021. "Endogenous piRNA-guided slicing triggers responder and trailer piRNA production from viral RNA in Aedes aegypti mosquitoes." Nucleic Acids Research , no. : 1.
PIWI-interacting (pi)RNAs are small silencing RNAs that are crucial for the defense against transposable elements in germline tissues of animals. In Aedes aegypti mosquitoes, the piRNA pathway also contributes to gene regulation in somatic tissues, illustrating additional roles for piRNAs and PIWI proteins besides transposon repression. Here, we identify a highly abundant endogenous piRNA (propiR1) that associates with both Piwi4 and Piwi5. PropiR1-mediated target silencing requires base pairing in the seed region with supplemental base pairing at the piRNA 3’ end. Yet, propiR1 represses a limited set of targets, among which the lncRNA AAEL027353 (lnc027353). Slicing of lnc027353 initiates production of responder and trailer piRNAs from the cleavage fragment. Expression of propiR1 commences early during embryonic development and mediates degradation of maternally provided lnc027353. Both propiR1 and its lncRNA target are conserved in the closely related Aedes albopictus mosquito, underscoring the importance of this regulatory network for mosquito development.
Valerie Betting; Joep Joosten; Rebecca Halbach; Melissa Thaler; Pascal Miesen; Ronald P Van Rij. A piRNA-lncRNA regulatory network initiates responder and trailer piRNA formation during mosquito embryonic development. RNA 2021, 1 .
AMA StyleValerie Betting, Joep Joosten, Rebecca Halbach, Melissa Thaler, Pascal Miesen, Ronald P Van Rij. A piRNA-lncRNA regulatory network initiates responder and trailer piRNA formation during mosquito embryonic development. RNA. 2021; ():1.
Chicago/Turabian StyleValerie Betting; Joep Joosten; Rebecca Halbach; Melissa Thaler; Pascal Miesen; Ronald P Van Rij. 2021. "A piRNA-lncRNA regulatory network initiates responder and trailer piRNA formation during mosquito embryonic development." RNA , no. : 1.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged as a new human pathogen in late 2019 and it has infected over 100 million people in less than a year. There is a clear need for effective antiviral drugs to complement current preventive measures, including vaccines. In this study, we demonstrate that berberine and obatoclax, two broad-spectrum antiviral compounds, are effective against multiple isolates of SARS-CoV-2. Berberine, a plant-derived alkaloid, inhibited SARS-CoV-2 at low micromolar concentrations and obatoclax, which was originally developed as an anti-apoptotic protein antagonist, was effective at sub-micromolar concentrations. Time-of-addition studies indicated that berberine acts on the late stage of the viral life cycle. In agreement, berberine mildly affected viral RNA synthesis, but it strongly reduced infectious viral titers, leading to an increase in the particle-to-pfu ratio. In contrast, obatoclax acted at the early stage of the infection, which is in line with its activity to neutralize the acidic environment in endosomes. We assessed infection of primary human nasal epithelial cells that were cultured on an air-liquid interface and found that SARS-CoV-2 infection induced and repressed expression of specific sets of cytokines and chemokines. Moreover, both obatoclax and berberine inhibited SARS-CoV-2 replication in these primary target cells. We propose berberine and obatoclax as potential antiviral drugs against SARS-CoV-2 that could be considered for further efficacy testing.
Finny Varghese; Esther van Woudenbergh; Gijs Overheul; Marc Eleveld; Lisa Kurver; Niels van Heerbeek; Arjan van Laarhoven; Pascal Miesen; Gerco Den Hartog; Marien de Jonge; Ronald van Rij. Berberine and Obatoclax Inhibit SARS-Cov-2 Replication in Primary Human Nasal Epithelial Cells In Vitro. Viruses 2021, 13, 282 .
AMA StyleFinny Varghese, Esther van Woudenbergh, Gijs Overheul, Marc Eleveld, Lisa Kurver, Niels van Heerbeek, Arjan van Laarhoven, Pascal Miesen, Gerco Den Hartog, Marien de Jonge, Ronald van Rij. Berberine and Obatoclax Inhibit SARS-Cov-2 Replication in Primary Human Nasal Epithelial Cells In Vitro. Viruses. 2021; 13 (2):282.
Chicago/Turabian StyleFinny Varghese; Esther van Woudenbergh; Gijs Overheul; Marc Eleveld; Lisa Kurver; Niels van Heerbeek; Arjan van Laarhoven; Pascal Miesen; Gerco Den Hartog; Marien de Jonge; Ronald van Rij. 2021. "Berberine and Obatoclax Inhibit SARS-Cov-2 Replication in Primary Human Nasal Epithelial Cells In Vitro." Viruses 13, no. 2: 282.
The genetic basis of antiviral immunity in dipteran insects is extensively studied in Drosophila melanogaster and advanced technologies for genetic manipulation allow a better characterization of immune responses also in non-model insect species. Especially, immunity in vector mosquitoes is recently in the spotlight, due to the medical impact that these insects have by transmitting viruses and other pathogens. Here, we review the current state of experimental evidence that supports antiviral functions for immune genes acting in different cellular pathways. We discuss the well-characterized RNA interference mechanism along with the less well-defined JAK-STAT, Toll, and IMD signaling pathways. Furthermore, we highlight the initial evidence for antiviral activity observed for the autophagy pathway, transcriptional pausing, as well as piRNA production from endogenous viral elements. We focus our review on studies from Drosophila and mosquito species from the lineages Aedes, Culex, and Anopheles, which contain major vector species responsible for virus transmission.
Samara Rosendo Machado; Tom van der Most; Pascal Miesen. Genetic determinants of antiviral immunity in dipteran insects – Compiling the experimental evidence. Developmental & Comparative Immunology 2021, 119, 104010 .
AMA StyleSamara Rosendo Machado, Tom van der Most, Pascal Miesen. Genetic determinants of antiviral immunity in dipteran insects – Compiling the experimental evidence. Developmental & Comparative Immunology. 2021; 119 ():104010.
Chicago/Turabian StyleSamara Rosendo Machado; Tom van der Most; Pascal Miesen. 2021. "Genetic determinants of antiviral immunity in dipteran insects – Compiling the experimental evidence." Developmental & Comparative Immunology 119, no. : 104010.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged as a new human pathogen in late 2019 and has infected an estimated 10% of the global population in less than a year. There is a clear need for effective antiviral drugs to complement current preventive measures including vaccines. In this study, we demonstrate that berberine and obatoclax, two broad-spectrum antiviral compounds, are effective against multiple isolates of SARS-CoV-2. Berberine, a plant-derived alkaloid, inhibited SARS-CoV-2 at low micromolar concentrations and obatoclax, originally developed as an anti-apoptotic protein antagonist, was effective at sub-micromolar concentrations. Time-of-addition studies indicated that berberine acts on the late stage of the viral life cycle. In agreement, berberine mildly affected viral RNA synthesis, but strongly reduced infectious viral titers, leading to an increase in the particle-to-pfu ratio. In contrast, obatoclax acted at the early stage of the infection, in line with its activity to neutralize the acidic environment in endosomes. We assessed infection of primary human nasal epithelial cells cultured on an air-liquid interface and found that SARS-CoV-2 infection induced and repressed expression of a specific set of cytokines and chemokines. Moreover, both obatoclax and berberine inhibited SARS-CoV-2 replication in these primary target cells. We propose berberine and obatoclax as potential antiviral drugs against SARS-CoV-2 that could be considered for further efficacy testing.
Finny S. Varghese; Esther van Woudenbergh; Gijs J. Overheul; Marc J. Eleveld; Lisa Kurver; Niels van Heerbeek; Arjan van Laarhoven; Pascal Miesen; Gerco Den Hartog; Marien I. de Jonge; Ronald P. van Rij. Berberine and obatoclax inhibit SARS-CoV-2 replication in primary human nasal epithelial cells in vitro. 2020, 1 .
AMA StyleFinny S. Varghese, Esther van Woudenbergh, Gijs J. Overheul, Marc J. Eleveld, Lisa Kurver, Niels van Heerbeek, Arjan van Laarhoven, Pascal Miesen, Gerco Den Hartog, Marien I. de Jonge, Ronald P. van Rij. Berberine and obatoclax inhibit SARS-CoV-2 replication in primary human nasal epithelial cells in vitro. . 2020; ():1.
Chicago/Turabian StyleFinny S. Varghese; Esther van Woudenbergh; Gijs J. Overheul; Marc J. Eleveld; Lisa Kurver; Niels van Heerbeek; Arjan van Laarhoven; Pascal Miesen; Gerco Den Hartog; Marien I. de Jonge; Ronald P. van Rij. 2020. "Berberine and obatoclax inhibit SARS-CoV-2 replication in primary human nasal epithelial cells in vitro." , no. : 1.
Summary Endogenous viral elements (EVEs) are viral sequences integrated in host genomes. A large number of non-retroviral EVEs was recently detected in Aedes mosquito genomes, leading to the hypothesis that mosquito EVEs may control exogenous infections by closely related viruses. Here, we experimentally investigated the role of an EVE naturally found in Aedes aegypti populations and derived from the widespread insect-specific virus, cell-fusing agent virus (CFAV). Using CRISPR-Cas9 genome editing, we created an Ae. aegypti line lacking the CFAV EVE. Absence of the EVE resulted in increased CFAV replication in ovaries, possibly modulating vertical transmission of the virus. Viral replication was controlled by targeting of viral RNA by EVE-derived P-element-induced wimpy testis-interacting RNAs (piRNAs). Our results provide evidence that antiviral piRNAs are produced in the presence of a naturally occurring EVE and its cognate virus, demonstrating a functional link between non-retroviral EVEs and antiviral immunity in a natural insect-virus interaction.
Yasutsugu Suzuki; Artem Baidaliuk; Pascal Miesen; Lionel Frangeul; Anna B. Crist; Sarah H. Merkling; Albin Fontaine; Sebastian Lequime; Isabelle Moltini-Conclois; Hervé Blanc; Ronald P. van Rij; Louis Lambrechts; Maria-Carla Saleh. Non-retroviral Endogenous Viral Element Limits Cognate Virus Replication in Aedes aegypti Ovaries. Current Biology 2020, 30, 3495 -3506.e6.
AMA StyleYasutsugu Suzuki, Artem Baidaliuk, Pascal Miesen, Lionel Frangeul, Anna B. Crist, Sarah H. Merkling, Albin Fontaine, Sebastian Lequime, Isabelle Moltini-Conclois, Hervé Blanc, Ronald P. van Rij, Louis Lambrechts, Maria-Carla Saleh. Non-retroviral Endogenous Viral Element Limits Cognate Virus Replication in Aedes aegypti Ovaries. Current Biology. 2020; 30 (18):3495-3506.e6.
Chicago/Turabian StyleYasutsugu Suzuki; Artem Baidaliuk; Pascal Miesen; Lionel Frangeul; Anna B. Crist; Sarah H. Merkling; Albin Fontaine; Sebastian Lequime; Isabelle Moltini-Conclois; Hervé Blanc; Ronald P. van Rij; Louis Lambrechts; Maria-Carla Saleh. 2020. "Non-retroviral Endogenous Viral Element Limits Cognate Virus Replication in Aedes aegypti Ovaries." Current Biology 30, no. 18: 3495-3506.e6.
In the germline of animals, PIWI interacting (pi)RNAs protect the genome against the detrimental effects of transposon mobilization. In Drosophila, piRNA-mediated cleavage of transposon RNA triggers the production of responder piRNAs via ping-pong amplification. Responder piRNA 3’ end formation is coupled to the production of downstream trailer piRNAs mediated by the nuclease Zucchini, expanding the repertoire of transposon piRNA sequences. In Aedes aegypti mosquitoes, piRNAs are generated from viral RNA, yet, it is unknown how viral piRNA 3’ ends are formed and whether viral RNA cleavage gives rise to trailer piRNA production. Here we report that in Ae. aegypti, virus- and transposon-derived piRNAs have sharp 3’ ends, and are biased for downstream uridine residues, features reminiscent of Zucchini cleavage of precursor piRNAs in Drosophila. We designed a reporter system to study viral piRNA 3’ end formation and found that targeting viral RNA by abundant endogenous piRNAs triggers the production of responder and trailer piRNAs. Using this reporter, we identified the Ae. aegypti orthologs of Zucchini and Nibbler, two nucleases involved in piRNA 3’ end formation. Our results furthermore suggest that autonomous piRNA production from viral RNA can be triggered and expanded by an initial cleavage event guided by genome-encoded piRNAs.
Joep Joosten; Gijs J. Overheul; Ronald P. Van Rij; Pascal Miesen. Endogenous piRNA-guided slicing triggers responder and trailer piRNA production from viral RNA in Aedes aegypti mosquitoes. 2020, 1 .
AMA StyleJoep Joosten, Gijs J. Overheul, Ronald P. Van Rij, Pascal Miesen. Endogenous piRNA-guided slicing triggers responder and trailer piRNA production from viral RNA in Aedes aegypti mosquitoes. . 2020; ():1.
Chicago/Turabian StyleJoep Joosten; Gijs J. Overheul; Ronald P. Van Rij; Pascal Miesen. 2020. "Endogenous piRNA-guided slicing triggers responder and trailer piRNA production from viral RNA in Aedes aegypti mosquitoes." , no. : 1.
SummaryEndogenous viral elements (EVEs) are viral sequences integrated in host genomes. A large number of non-retroviral EVEs was recently detected in Aedes mosquito genomes, leading to the hypothesis that mosquito EVEs may control exogenous infections by closely related viruses. Here, we experimentally investigated the role of an EVE naturally found in Aedes aegypti populations and derived from the widespread insect-specific virus, cell-fusing agent virus (CFAV). Using CRISPR/Cas9 genome editing, we created an Ae. aegypti line lacking the CFAV EVE. Absence of the EVE resulted in increased CFAV replication in ovaries, possibly modulating vertical transmission of the virus. Viral replication was controlled by targeting of viral RNA by EVE-derived piRNAs. Our results provide evidence that antiviral piRNAs are produced in the presence of a naturally occurring EVE and its cognate virus, demonstrating a functional link between non-retroviral EVEs and antiviral immunity in a natural insect-virus interaction.
Yasutsugu Suzuki; Artem Baidaliuk; Pascal Miesen; Lionel Frangeul; Anna B. Crist; Sarah H. Merkling; Albin Fontaine; Sebastian Lequime; Isabelle Moltini-Conclois; Hervé Blanc; Ronald P. Van Rij; Louis Lambrechts; Maria-Carla Saleh. Non-retroviral endogenous viral element limits cognate virus replication in Aedes aegypti ovaries. 2020, 1 .
AMA StyleYasutsugu Suzuki, Artem Baidaliuk, Pascal Miesen, Lionel Frangeul, Anna B. Crist, Sarah H. Merkling, Albin Fontaine, Sebastian Lequime, Isabelle Moltini-Conclois, Hervé Blanc, Ronald P. Van Rij, Louis Lambrechts, Maria-Carla Saleh. Non-retroviral endogenous viral element limits cognate virus replication in Aedes aegypti ovaries. . 2020; ():1.
Chicago/Turabian StyleYasutsugu Suzuki; Artem Baidaliuk; Pascal Miesen; Lionel Frangeul; Anna B. Crist; Sarah H. Merkling; Albin Fontaine; Sebastian Lequime; Isabelle Moltini-Conclois; Hervé Blanc; Ronald P. Van Rij; Louis Lambrechts; Maria-Carla Saleh. 2020. "Non-retroviral endogenous viral element limits cognate virus replication in Aedes aegypti ovaries." , no. : 1.
PIWI-interacting (pi)RNAs are small silencing RNAs that are crucial for the defense against transposable elements in germline tissues of animals. In Aedes aegypti mosquitoes, the piRNA pathway also contributes to gene regulation in somatic tissues, illustrating additional roles for piRNAs and PIWI proteins besides transposon repression. Here, we identify a highly abundant endogenous piRNA (propiR1) that associates with both Piwi4 and Piwi5. PropiR1-mediated target silencing requires base pairing in the seed region with supplemental base pairing at the piRNA 3’ end. Yet, propiR1 represses a limited set of targets, among which the lncRNA AAEL027353 (lnc027353). Slicing of lnc027353 initiates production of responder and trailer piRNAs from the cleavage fragment. Expression of propiR1 commences early during embryonic development and mediates degradation of maternally provided lnc027353. Both propiR1 and its lncRNA target are conserved in the closely related Aedes albopictus mosquito, underscoring the importance of this regulatory network for mosquito development.
Valerie Betting; Joep Joosten; Rebecca Halbach; Melissa Thaler; Pascal Miesen; Ronald P. Van Rij. A piRNA-lncRNA regulatory network initiates responder and trailer piRNA formation during mosquito embryonic development. 2020, 1 .
AMA StyleValerie Betting, Joep Joosten, Rebecca Halbach, Melissa Thaler, Pascal Miesen, Ronald P. Van Rij. A piRNA-lncRNA regulatory network initiates responder and trailer piRNA formation during mosquito embryonic development. . 2020; ():1.
Chicago/Turabian StyleValerie Betting; Joep Joosten; Rebecca Halbach; Melissa Thaler; Pascal Miesen; Ronald P. Van Rij. 2020. "A piRNA-lncRNA regulatory network initiates responder and trailer piRNA formation during mosquito embryonic development." , no. : 1.
The Asian tiger mosquito Aedes albopictus is globally expanding and has become the main vector for human arboviruses in Europe. Here we present AalbF2, a dramatically improved assembly of the Ae. albopictus genome that has revealed widespread viral insertions, novel microRNAs and piRNA clusters, the sex determining locus, new immunity genes, and has enabled genome-wide studies of geographically diverse Ae. albopictus populations and analyses of the developmental and stage-dependent network of expression data. Additionally, we built the first physical map for this species with 75% of the assembled genome anchored to the chromosomes. These up-to-date resources of the genome provide a foundation to improve understanding of the adaptation potential and the epidemiological relevance of this species and foster the development of innovative control measures.One Sentence SummaryLong-read and Hi-C-based de novo assembly of the arboviral vector Aedes albopictus genome fosters deeper understanding of its biological features.
Umberto Palatini; R.A. Masri; Luciano V Cosme; Sergey Koren; Francoise Thibaud-Nissen; James K Biedler; Flavia Krsticevic; Spencer Johnston; Rebecca Halbach; Jacob E Crawford; Igor Antoshechkin; Anna-Bella Failloux; Elisa Pischedda; Michele Marconcini; Jay Ghurye; Arang Rhie; Atashi Sharma; Dmitrii A Karagodin; J. Jenrette; Stephanie Gamez; Pascal Miesen; Adalgisa Caccone; Maria V Sharakhova; Zhijian Tu; Philippos Aris Papathanos; Ronald P. Van Rij; Omar S Akbari; Jeffrey Powell; Adam M. Phillippy; Bonizzoni M.. Improved reference genome of the arboviral vector Aedes albopictus. 2020, 1 .
AMA StyleUmberto Palatini, R.A. Masri, Luciano V Cosme, Sergey Koren, Francoise Thibaud-Nissen, James K Biedler, Flavia Krsticevic, Spencer Johnston, Rebecca Halbach, Jacob E Crawford, Igor Antoshechkin, Anna-Bella Failloux, Elisa Pischedda, Michele Marconcini, Jay Ghurye, Arang Rhie, Atashi Sharma, Dmitrii A Karagodin, J. Jenrette, Stephanie Gamez, Pascal Miesen, Adalgisa Caccone, Maria V Sharakhova, Zhijian Tu, Philippos Aris Papathanos, Ronald P. Van Rij, Omar S Akbari, Jeffrey Powell, Adam M. Phillippy, Bonizzoni M.. Improved reference genome of the arboviral vector Aedes albopictus. . 2020; ():1.
Chicago/Turabian StyleUmberto Palatini; R.A. Masri; Luciano V Cosme; Sergey Koren; Francoise Thibaud-Nissen; James K Biedler; Flavia Krsticevic; Spencer Johnston; Rebecca Halbach; Jacob E Crawford; Igor Antoshechkin; Anna-Bella Failloux; Elisa Pischedda; Michele Marconcini; Jay Ghurye; Arang Rhie; Atashi Sharma; Dmitrii A Karagodin; J. Jenrette; Stephanie Gamez; Pascal Miesen; Adalgisa Caccone; Maria V Sharakhova; Zhijian Tu; Philippos Aris Papathanos; Ronald P. Van Rij; Omar S Akbari; Jeffrey Powell; Adam M. Phillippy; Bonizzoni M.. 2020. "Improved reference genome of the arboviral vector Aedes albopictus." , no. : 1.
Tandem repeat elements such as the highly diverse class of satellite repeats occupy large parts of eukaryotic chromosomes. Most occur at (peri)centromeric and (sub)telomeric regions and have been implicated in chromosome organization, stabilization, and segregation1. Others are located more dispersed throughout the genome, but their functions remained largely enigmatic. Satellite repeats in euchromatic regions were hypothesized to regulate gene expression in cis by modulation of the local heterochromatin, or in trans via repeat-derived transcripts2,3. Yet, due to a lack of experimental models, gene regulatory potential of satellite repeats remains largely unexplored. Here we show that, in the vector mosquito Aedes aegypti, a satellite repeat promotes sequence-specific gene silencing via the expression of two abundant PIWI-interacting RNAs (piRNAs). Strikingly, whereas satellite repeats and piRNA sequences generally evolve extremely fast4-6, this locus was conserved for approximately 200 million years, suggesting a central function in mosquito biology. Tandem repeat-derived piRNA production commenced shortly after egg-laying and inactivation of the most abundant of the two piRNAs in early embryos resulted in an arrest of embryonic development. Transcriptional profiling in these embryos revealed the failure to degrade maternally provided transcripts that are normally cleared during maternal-to-zygotic transition. Our results reveal a novel mechanism in which satellite repeats regulate global gene expression in trans via piRNA-mediated gene silencing, which is fundamental to embryonic development. These findings highlight the regulatory potential of this enigmatic class of repeats.
Rebecca Halbach; Pascal Miesen; Joep Joosten; Ezgi Taşköprü; Bas Pennings; Chantal B.F. Vogels; Sarah H. Merkling; Constantianus J. Koenraadt; Louis Lambrechts; Ronald P. Van Rij. An ancient satellite repeat controls gene expression and embryonic development in Aedes aegypti through a highly conserved piRNA. 2020, 1 .
AMA StyleRebecca Halbach, Pascal Miesen, Joep Joosten, Ezgi Taşköprü, Bas Pennings, Chantal B.F. Vogels, Sarah H. Merkling, Constantianus J. Koenraadt, Louis Lambrechts, Ronald P. Van Rij. An ancient satellite repeat controls gene expression and embryonic development in Aedes aegypti through a highly conserved piRNA. . 2020; ():1.
Chicago/Turabian StyleRebecca Halbach; Pascal Miesen; Joep Joosten; Ezgi Taşköprü; Bas Pennings; Chantal B.F. Vogels; Sarah H. Merkling; Constantianus J. Koenraadt; Louis Lambrechts; Ronald P. Van Rij. 2020. "An ancient satellite repeat controls gene expression and embryonic development in Aedes aegypti through a highly conserved piRNA." , no. : 1.
Coevolution of viruses and their hosts may lead to viral strategies to avoid, evade, or suppress antiviral immunity. An example is antiviral RNA interference (RNAi) in insects: the host RNAi machinery processes viral double-stranded RNA into small interfering RNAs (siRNAs) to suppress viral replication, whereas insect viruses encode suppressors of RNAi, many of which inhibit viral small interfering RNA (vsiRNA) production. Yet, many studies have analyzed viral RNAi suppressors in heterologous systems, due to the lack of experimental systems to manipulate the viral genome of interest, raising questions about in vivo functions of RNAi suppressors. To address this caveat, we generated an RNAi suppressor-defective mutant of invertebrate iridescent virus 6 (IIV6), a large DNA virus in which we previously identified the 340R protein as a suppressor of RNAi. Loss of 340R did not affect vsiRNA production, indicating that 340R binds siRNA duplexes to prevent RNA-induced silencing complex assembly. Indeed, vsiRNAs were not efficiently loaded into Argonaute 2 during wild-type IIV6 infection. Moreover, IIV6 induced a limited set of mature microRNAs in a 340R-dependent manner, most notably miR-305–3p, which we attribute to stabilization of the miR-305–5p:3p duplex by 340R. The IIV6 340R deletion mutant did not have a replication defect in cells, but was strongly attenuated in adult Drosophila. This in vivo replication defect was completely rescued in RNAi mutant flies, indicating that 340R is a bona fide RNAi suppressor, the absence of which uncovers a potent antiviral immune response that suppresses virus accumulation ∼100-fold. Together, our work indicates that viral RNAi suppressors may completely mask antiviral immunity.
Alfred W. Bronkhorst; Rob Vogels; Gijs J. Overheul; Bas Pennings; Valérie Gausson-Dorey; Pascal Miesen; Ronald P. van Rij. A DNA virus-encoded immune antagonist fully masks the potent antiviral activity of RNAi in Drosophila. Proceedings of the National Academy of Sciences 2019, 116, 24296 -24302.
AMA StyleAlfred W. Bronkhorst, Rob Vogels, Gijs J. Overheul, Bas Pennings, Valérie Gausson-Dorey, Pascal Miesen, Ronald P. van Rij. A DNA virus-encoded immune antagonist fully masks the potent antiviral activity of RNAi in Drosophila. Proceedings of the National Academy of Sciences. 2019; 116 (48):24296-24302.
Chicago/Turabian StyleAlfred W. Bronkhorst; Rob Vogels; Gijs J. Overheul; Bas Pennings; Valérie Gausson-Dorey; Pascal Miesen; Ronald P. van Rij. 2019. "A DNA virus-encoded immune antagonist fully masks the potent antiviral activity of RNAi in Drosophila." Proceedings of the National Academy of Sciences 116, no. 48: 24296-24302.
The RNA interference (RNAi) pathway is a potent antiviral defense mechanism in plants and invertebrates, in response to which viruses evolved suppressors of RNAi. In mammals, the first line of defense is mediated by the type I interferon system (IFN); however, the degree to which RNAi contributes to antiviral defense is still not completely understood. Recent work suggests that antiviral RNAi is active in undifferentiated stem cells and that antiviral RNAi can be uncovered in differentiated cells in which the IFN system is inactive or in infections with viruses lacking putative viral suppressors of RNAi. In this review, we describe the mechanism of RNAi and its antiviral functions in insects and mammals. We draw parallels and highlight differences between (antiviral) RNAi in these classes of animals and discuss open questions for future research.
Susan Schuster; Pascal Miesen; Ronald P. Van Rij. Antiviral RNAi in Insects and Mammals: Parallels and Differences. Viruses 2019, 11, 448 .
AMA StyleSusan Schuster, Pascal Miesen, Ronald P. Van Rij. Antiviral RNAi in Insects and Mammals: Parallels and Differences. Viruses. 2019; 11 (5):448.
Chicago/Turabian StyleSusan Schuster; Pascal Miesen; Ronald P. Van Rij. 2019. "Antiviral RNAi in Insects and Mammals: Parallels and Differences." Viruses 11, no. 5: 448.
Small RNA mediated responses are essential for antiviral defence in mosquitoes, however, they appear to differ per virus-vector combination. To further investigate the diversity of small RNA responses against viruses in mosquitoes, we applied a small RNA deep sequencing approach on five mosquito cell lines: Culex tarsalis CT cells, Aedes albopictus U4.4 and C6/36 cells, Ae. aegypti Aag2 cells (cleared from cell fusing agent virus and Culex Y virus (CYV) by repetitive dsRNA transfections) and Ae. pseudoscutellaris AP-61 cells. De novo assembly of small RNAs revealed the presence of Phasi Charoen-like virus (PCLV), Calbertado virus, Flock House virus and a novel narnavirus in CT cells, CYV in U4.4 cells, and PCLV in Aag2 cells, whereas no insect-specific viruses (ISVs) were detected in C6/36 and AP-61 cells. Next, we investigated the small RNA responses to the identified ISVs and to acute infection with the arthropod-borne West Nile virus (WNV). We demonstrate that AP-61 and C6/36 cells do not produce siRNAs to WNV infection, suggesting that AP-61, like C6/36, are Dicer-2 deficient. CT cells produced a strong siRNA response to the persistent ISVs and acute WNV infection. Interestingly, CT cells also produced viral PIWI-interacting (pi)RNAs to PCLV, but not to WNV or any of the other ISVs. In contrast, in U4.4 and Aag2 cells, WNV siRNAs, and pi-like RNAs without typical ping-pong piRNA signature were observed, while this signature was present in PCLV piRNAs in Aag2 cells. Together, our results demonstrate that mosquito small RNA responses are strongly dependent on both the mosquito cell type and/or the mosquito species and family of the infecting virus.
Giel P. Göertz; Pascal Miesen; Gijs J. Overheul; Ronald P. Van Rij; Monique M. Van Oers; Gorben P. Pijlman. Mosquito Small RNA Responses to West Nile and Insect-Specific Virus Infections in Aedes and Culex Mosquito Cells. Viruses 2019, 11, 271 .
AMA StyleGiel P. Göertz, Pascal Miesen, Gijs J. Overheul, Ronald P. Van Rij, Monique M. Van Oers, Gorben P. Pijlman. Mosquito Small RNA Responses to West Nile and Insect-Specific Virus Infections in Aedes and Culex Mosquito Cells. Viruses. 2019; 11 (3):271.
Chicago/Turabian StyleGiel P. Göertz; Pascal Miesen; Gijs J. Overheul; Ronald P. Van Rij; Monique M. Van Oers; Gorben P. Pijlman. 2019. "Mosquito Small RNA Responses to West Nile and Insect-Specific Virus Infections in Aedes and Culex Mosquito Cells." Viruses 11, no. 3: 271.
Commensal bacteria that colonize the midgut of Aedes aegypti mosquitoes can influence the transmission of arthropod-borne viruses. In this issue of Cell Host & Microbe, Wu et al. (2019) show that Serratia marcescens bacteria secrete enhancin proteins that cleave membrane-bound mucins, thereby facilitating dengue virus infection of midgut epithelial cells.
Pascal Miesen; Ronald P. Van Rij. Crossing the Mucosal Barrier: A Commensal Bacterium Gives Dengue Virus a Leg-Up in the Mosquito Midgut. Cell Host & Microbe 2019, 25, 1 -2.
AMA StylePascal Miesen, Ronald P. Van Rij. Crossing the Mucosal Barrier: A Commensal Bacterium Gives Dengue Virus a Leg-Up in the Mosquito Midgut. Cell Host & Microbe. 2019; 25 (1):1-2.
Chicago/Turabian StylePascal Miesen; Ronald P. Van Rij. 2019. "Crossing the Mucosal Barrier: A Commensal Bacterium Gives Dengue Virus a Leg-Up in the Mosquito Midgut." Cell Host & Microbe 25, no. 1: 1-2.
PIWI-interacting RNAs (piRNAs) comprise a class of small RNAs best known for suppressing transposable elements in germline tissues. The vector mosquito Aedes aegypti encodes seven PIWI genes, four of which are somatically expressed. This somatic piRNA pathway generates piRNAs from viral RNA during infection with cytoplasmic RNA viruses through ping-pong amplification by the PIWI proteins Ago3 and Piwi5. Yet, additional insights into the molecular mechanisms mediating non-canonical piRNA production are lacking. TUDOR-domain containing (Tudor) proteins facilitate piRNA biogenesis in Drosophila melanogaster and other model organisms. We thus hypothesized that Tudor proteins are required for viral piRNA production and performed a knockdown screen targeting all A. aegypti Tudor genes. Knockdown of the Tudor genes AAEL012437, Vreteno, Yb, SMN and AAEL008101-RB resulted in significantly reduced viral piRNA levels, with AAEL012437-depletion having the strongest effect. This protein, which we named Veneno, associates directly with Ago3 in an sDMA-dependent manner and localizes in cytoplasmic foci reminiscent of piRNA processing granules of Drosophila. Veneno-interactome analyses reveal a network of co-factors including the orthologs of the Drosophila piRNA pathway components Vasa and Yb, which in turn interacts with Piwi5. We propose that Veneno assembles a multi-protein complex for ping-pong dependent piRNA production from viral RNA.
Joep Joosten; Pascal Miesen; Ezgi Taşköprü; Bas Pennings; Pascal W T C Jansen; Martijn A Huynen; Michiel Vermeulen; Ronald P Van Rij. The Tudor protein Veneno assembles the ping-pong amplification complex that produces viral piRNAs in Aedes mosquitoes. Nucleic Acids Research 2018, 47, 2546 -2559.
AMA StyleJoep Joosten, Pascal Miesen, Ezgi Taşköprü, Bas Pennings, Pascal W T C Jansen, Martijn A Huynen, Michiel Vermeulen, Ronald P Van Rij. The Tudor protein Veneno assembles the ping-pong amplification complex that produces viral piRNAs in Aedes mosquitoes. Nucleic Acids Research. 2018; 47 (5):2546-2559.
Chicago/Turabian StyleJoep Joosten; Pascal Miesen; Ezgi Taşköprü; Bas Pennings; Pascal W T C Jansen; Martijn A Huynen; Michiel Vermeulen; Ronald P Van Rij. 2018. "The Tudor protein Veneno assembles the ping-pong amplification complex that produces viral piRNAs in Aedes mosquitoes." Nucleic Acids Research 47, no. 5: 2546-2559.
SummaryTUDOR-domain containing proteins facilitate PIWI interacting (pi)RNA biogenesis in Drosophila melanogaster and other model organisms. In Aedes aegypti mosquitoes, a somatically active piRNA pathway generates piRNAs from viral RNA during acute infection with cytoplasmic RNA viruses. Viral piRNA biogenesis requires ping-pong amplification by the PIWI proteins Ago3 and Piwi5. We hypothesized that Tudor proteins are required for viral piRNA production and performed a knockdown screen targeting all Ae. aegypti Tudor genes. Knockdown of several Tudor genes resulted in reduced viral piRNA levels, with silencing of AAEL012437 having the strongest effect. This protein, which we named Veneno, associates directly with Ago3 in an sDMA-dependent manner and localizes in cytoplasmic foci reminiscent of piRNA processing granules of Drosophila. Veneno-interactome analyses reveal a network of co-factors including the orthologs of the Drosophila piRNA pathway components Vasa and Yb, which in turn interacts directly with Piwi5. We propose that Veneno assembles a multi-protein complex for ping-pong dependent piRNA production from exogenous viral RNA.
Joep Joosten; Pascal Miesen; Bas Pennings; Pascal W.T.C. Jansen; Martijn A. Huynen; Michiel Vermeulen; Ronald P. Van Rij. The Tudor protein Veneno assembles the ping-pong amplification complex that 1 produces viral piRNAs in Aedes mosquitoes. 2018, 242305 .
AMA StyleJoep Joosten, Pascal Miesen, Bas Pennings, Pascal W.T.C. Jansen, Martijn A. Huynen, Michiel Vermeulen, Ronald P. Van Rij. The Tudor protein Veneno assembles the ping-pong amplification complex that 1 produces viral piRNAs in Aedes mosquitoes. . 2018; ():242305.
Chicago/Turabian StyleJoep Joosten; Pascal Miesen; Bas Pennings; Pascal W.T.C. Jansen; Martijn A. Huynen; Michiel Vermeulen; Ronald P. Van Rij. 2018. "The Tudor protein Veneno assembles the ping-pong amplification complex that 1 produces viral piRNAs in Aedes mosquitoes." , no. : 242305.
Arthropod-borne viruses (arboviruses) transmitted by mosquito vectors cause many important emerging or resurging infectious diseases in humans including dengue, chikungunya and Zika. Understanding the co-evolutionary processes among viruses and vectors is essential for the development of novel transmission-blocking strategies. Episomal viral DNA fragments are produced from arboviral RNA upon infection of mosquito cells and adults. Additionally, sequences from insect-specific viruses and arboviruses have been found integrated into mosquito genomes. We used a bioinformatic approach to analyse the presence, abundance, distribution, and transcriptional activity of integrations from 425 non-retroviral viruses, including 133 arboviruses, across the presently available 22 mosquito genome sequences. Large differences in abundance and types of viral integrations were observed in mosquito species from the same region. Viral integrations are unexpectedly abundant in the arboviral vector species Aedes aegypti and Ae. albopictus, in which they are approximately ~10-fold more abundant than in other mosquito species analysed. Additionally, viral integrations are enriched in piRNA clusters of both the Ae. aegypti and Ae. albopictus genomes and, accordingly, they express piRNAs, but not siRNAs. Differences in the number of viral integrations in the genomes of mosquito species from the same geographic area support the conclusion that integrations of viral sequences is not dependent on viral exposure, but that lineage-specific interactions exist. Viral integrations are abundant in Ae. aegypti and Ae. albopictus, and represent a thus far underappreciated component of their genomes. Additionally, the genome locations of viral integrations and their production of piRNAs indicate a functional link between viral integrations and the piRNA pathway. These results greatly expand the breadth and complexity of small RNA-mediated regulation and suggest a role for viral integrations in antiviral defense in these two mosquito species.
Umberto Palatini; Pascal Miesen; Rebeca Carballar-Lejarazu; Lino Ometto; Ettore Rizzo; Zhijian Tu; Ronald P. van Rij; Mariangela Bonizzoni. Comparative genomics shows that viral integrations are abundant and express piRNAs in the arboviral vectors Aedes aegypti and Aedes albopictus. BMC Genomics 2017, 18, 1 -15.
AMA StyleUmberto Palatini, Pascal Miesen, Rebeca Carballar-Lejarazu, Lino Ometto, Ettore Rizzo, Zhijian Tu, Ronald P. van Rij, Mariangela Bonizzoni. Comparative genomics shows that viral integrations are abundant and express piRNAs in the arboviral vectors Aedes aegypti and Aedes albopictus. BMC Genomics. 2017; 18 (1):1-15.
Chicago/Turabian StyleUmberto Palatini; Pascal Miesen; Rebeca Carballar-Lejarazu; Lino Ometto; Ettore Rizzo; Zhijian Tu; Ronald P. van Rij; Mariangela Bonizzoni. 2017. "Comparative genomics shows that viral integrations are abundant and express piRNAs in the arboviral vectors Aedes aegypti and Aedes albopictus." BMC Genomics 18, no. 1: 1-15.
BackgroundArthropod-borne viruses (arboviruses) transmitted by mosquito vectors cause many important emerging or resurging infectious diseases in humans including dengue, chikungunya and Zika. Understanding the co-evolutionary processes among viruses and vectors is essential for the development of novel transmission-blocking strategies. Arboviruses form episomal viral DNA fragments upon infection of mosquito cells and adults. Additionally, sequences from insect-specific viruses and arboviruses have been found integrated into mosquito genomes.ResultsWe used a bioinformatic approach to analyze the presence, abundance, distribution, and transcriptional activity of integrations from 425 non-retroviral viruses, including 133 arboviruses, across the presently available 22 mosquito genome sequences. Large differences in abundance and types of viral integrations were observed in mosquito species from the same region. Viral integrations are unexpectedly abundant in the arboviral vector species Aedes aegypti and Ae. albopictus, but are ∼10-fold less abundant in all other mosquitoes analysed. Additionally, viral integrations are enriched in piRNA clusters of both the Ae. aegypti and Ae. albopictus genomes and, accordingly, they express piRNAs, but not siRNAs.ConclusionsDifferences in number of viral integrations in the genomes of mosquito species from the same geographic area support the conclusion that integrations of viral sequences is not dependent on viral exposure, but that lineage-specific interactions exits. Viral integrations are abundant in Ae. aegypti and Ae. albopictus, and represent a thus far unappreciated component of their genomes. Additionally, the genome locations of viral integrations and their production of piRNAs indicate a functional link between viral integrations and the piRNA pathway. These results greatly expand the breadth and complexity of small RNA-mediated regulation and suggest a role for viral integrations in antiviral defense in these two mosquito species.
Umberto Palatini; Pascal Miesen; Rebeca Carballar-Lejarazu; Lino Ometto; Ettore Rizzo; Zhijian Tu; Ronald Van Rij; Mariangela Bonizzoni. Comparative genomics shows that viral integrations are abundant and express piRNAs in the arboviral vectors Aedes aegypti and Aedes albopictus. 2017, 128637 .
AMA StyleUmberto Palatini, Pascal Miesen, Rebeca Carballar-Lejarazu, Lino Ometto, Ettore Rizzo, Zhijian Tu, Ronald Van Rij, Mariangela Bonizzoni. Comparative genomics shows that viral integrations are abundant and express piRNAs in the arboviral vectors Aedes aegypti and Aedes albopictus. . 2017; ():128637.
Chicago/Turabian StyleUmberto Palatini; Pascal Miesen; Rebeca Carballar-Lejarazu; Lino Ometto; Ettore Rizzo; Zhijian Tu; Ronald Van Rij; Mariangela Bonizzoni. 2017. "Comparative genomics shows that viral integrations are abundant and express piRNAs in the arboviral vectors Aedes aegypti and Aedes albopictus." , no. : 128637.
The piRNA pathway is of key importance in controlling transposable elements in most animal species. In the vector mosquito Aedes aegypti, the presence of eight PIWI proteins and the accumulation of viral piRNAs upon arbovirus infection suggest additional functions of the piRNA pathway beyond genome defense. To better understand the regulatory potential of this pathway, we analyzed in detail host-derived piRNAs in A. aegypti Aag2 cells. We show that a large repertoire of protein-coding genes and non-retroviral integrated RNA virus elements are processed into genic piRNAs by different combinations of PIWI proteins. Among these, we identify a class of genes that produces piRNAs from coding sequences in an Ago3- and Piwi5-dependent fashion. We demonstrate that the replication-dependent histone gene family is a genic source of ping-pong dependent piRNAs and that histone-derived piRNAs are dynamically expressed throughout the cell cycle, suggesting a role for the piRNA pathway in the regulation of histone gene expression. Moreover, our results establish the Aag2 cell line as an accessible experimental model to study gene-derived piRNAs.
Erika Girardi; Pascal Miesen; Bas Pennings; Lionel Frangeul; Maria-Carla Saleh; Ronald P. Van Rij. Histone-derived piRNA biogenesis depends on the ping-pong partners Piwi5 and Ago3 inAedes aegypti. Nucleic Acids Research 2017, 45, 4881 -4892.
AMA StyleErika Girardi, Pascal Miesen, Bas Pennings, Lionel Frangeul, Maria-Carla Saleh, Ronald P. Van Rij. Histone-derived piRNA biogenesis depends on the ping-pong partners Piwi5 and Ago3 inAedes aegypti. Nucleic Acids Research. 2017; 45 (8):4881-4892.
Chicago/Turabian StyleErika Girardi; Pascal Miesen; Bas Pennings; Lionel Frangeul; Maria-Carla Saleh; Ronald P. Van Rij. 2017. "Histone-derived piRNA biogenesis depends on the ping-pong partners Piwi5 and Ago3 inAedes aegypti." Nucleic Acids Research 45, no. 8: 4881-4892.