Sequences for Lloviu virus (LLOV), a putative novel filovirus, were first identified in Miniopterus schreibersii bats in Spain following a massive bat die-off in 2002, and also recently found in bats in Hungary. However, until now it is unclear if these sequences correspond to a fully functional, infectious virus, and whether it will show a pathogenic phenotype like African filoviruses, such as ebola- and marburgviruses, or be apathogenic for humans, like the Asian filovirus Reston virus. Since no infectious virus has been recovered, the only opportunity to study infectious LLOV is to use a reverse genetics-based full-length clone system to de novo generate LLOV. As a first step in this process, and to investigate whether the identified sequences indeed correspond to functional viral proteins, we have developed life cycle modelling systems for LLOV, which allow us to study genome replication and transcription as well as entry of this virus. We show that all LLOV proteins fulfill their canonical role in the virus life cycle as expected based on the well-studied related filovirus Ebola virus. Further, we have analysed the intergenus-compatibility of proteins that have to act in concert to facilitate the virus life cycle. We show that some but not all proteins from LLOV and Ebola virus are compatible with each other, emphasizing the close relationship of these viruses, and informing future studies of filovirus biology with respect to the generation of genus-chimeric proteins in order to probe virus protein–protein interactions on a functional level.
GrosethA, MarziA, HoenenT, HerwigA, GardnerD et al. The Ebola virus glycoprotein contributes to but is not sufficient for virulence in vivo. PLoS Pathog2012; 8:e1002847 [View Article]
NegredoA, PalaciosG, Vázquez-MorónS, GonzálezF, DopazoH et al. Discovery of an ebolavirus-like filovirus in Europe. PLoS Pathog2011; 7:e1002304 [View Article]
BharatTAM, NodaT, RichesJD, KraehlingV, KolesnikovaL et al. Structural dissection of Ebola virus and its assembly determinants using cryo-electron tomography. Proc Natl Acad Sci U S A2012; 109:4275–4280 [View Article]
BharatTAM, RichesJD, KolesnikovaL, WelschS, KrählingV et al. Cryo-electron tomography of Marburg virus particles and their morphogenesis within infected cells. PLoS Biol2011; 9:e1001196 [View Article]
BeniacDR, MelitoPL, DevarennesSL, HiebertSL, RabbMJ et al. The organisation of Ebola virus reveals a capacity for extensive, modular polyploidy. PLoS One2012; 7:e29608 [View Article]
MühlbergerE, LötferingB, KlenkHD, BeckerS. Three of the four nucleocapsid proteins of Marburg virus, NP, VP35, and L, are sufficient to mediate replication and transcription of Marburg virus-specific monocistronic minigenomes. J Virol1998; 72:8756–8764
MühlbergerE, WeikM, VolchkovVE, KlenkHD, BeckerS. Comparison of the transcription and replication strategies of Marburg virus and Ebola virus by using artificial replication systems. J Virol1999; 73:2333–2342
EnterleinS, VolchkovV, WeikM, KolesnikovaL, VolchkovaV et al. Rescue of recombinant Marburg virus from cDNA is dependent on nucleocapsid protein VP30. J Virol2006; 80:1038–1043 [View Article]
HoenenT, JungS, HerwigA, GrosethA, BeckerS. Both matrix proteins of Ebola virus contribute to the regulation of viral genome replication and transcription. Virology2010; 403:56–66 [View Article]
WattA, MoukambiF, BanadygaL, GrosethA, CallisonJ et al. A novel life cycle modeling system for Ebola virus shows a genome length-dependent role of VP24 in virus infectivity. J Virol2014; 88:10511–10524 [View Article]
JasenoskyLD, NeumannG, LukashevichI, KawaokaY. Ebola virus VP40-induced particle formation and association with the lipid bilayer. J Virol2001; 75:5205–5214 [View Article]
SwensonDL, WarfieldKL, KuehlK, LarsenT, HeveyMC et al. Generation of Marburg virus-like particles by co-expression of glycoprotein and matrix protein. FEMS Immunol Med Microbiol2004; 40:27–31 [View Article]
NanboA, ImaiM, WatanabeS, NodaT, TakahashiK et al. Ebolavirus is internalized into host cells via macropinocytosis in a viral glycoprotein-dependent manner. PLoS Pathog2010; 6:e1001121 [View Article]
KondratowiczAS, LennemannNJ, SinnPL, DaveyRA, HuntCL et al. T-cell immunoglobulin and mucin domain 1 (Tim-1) is a receptor for Zaire ebolavirus and Lake Victoria marburgvirus. Proc Natl Acad Sci U S A2011; 108:8426–8431 [View Article]
HartyRN, BrownME, WangG, HuibregtseJ, HayesFP. A PPxY motif within the VP40 protein of Ebola virus interacts physically and functionally with a ubiquitin ligase: implications for filovirus budding. Proc Natl Acad Sci U S A2000; 97:13871–13876 [View Article]
LicataJM, Simpson-HolleyM, WrightNT, HanZ, ParagasJ et al. Overlapping motifs (PTAP and PPEY) within the Ebola virus VP40 protein function independently as late budding domains: involvement of host proteins TSG101 and VPS-4. J Virol2003; 77:1812–1819 [View Article]
Martin-SerranoJ, ZangT, BieniaszPD. HIV-1 and Ebola virus encode small peptide motifs that recruit TSG101 to sites of particle assembly to facilitate egress. Nat Med2001; 7:1313–1319 [View Article]
BrinkmannC, NehlmeierI, Walendy-GnirßK, NehlsJ, González HernándezM et al. The tetherin antagonism of the Ebola virus glycoprotein requires an intact receptor-binding domain and can be blocked by GP1-Specific antibodies. J Virol2016; 90:11075–11086 [View Article]
MaruyamaJ, MiyamotoH, KajiharaM, OgawaH, MaedaK et al. Characterization of the envelope glycoprotein of a novel filovirus, lloviu virus. J Virol2014; 88:99–109 [View Article]
FeaginsAR, BaslerCF. Lloviu virus VP24 and VP35 proteins function as innate immune antagonists in human and bat cells. Virology2015; 485:145–152 [View Article]
HoenenT, GrosethA, de Kok-MercadoF, KuhnJH, Wahl-JensenV. Minigenomes, transcription and replication competent virus-like particles and beyond: reverse genetics systems for filoviruses and other negative stranded hemorrhagic fever viruses. Antiviral Res2011; 91:195–208 [View Article]
VolchkovVE, VolchkovaVA, ChepurnovAA, BlinovVM, DolnikO et al. Characterization of the L gene and 5' trailer region of Ebola virus. J Gen Virol1999; 80:355–362 [View Article]
NeumannG, EbiharaH, TakadaA, NodaT, KobasaD et al. Ebola virus VP40 late domains are not essential for viral replication in cell culture. J Virol2005; 79:10300–10307 [View Article]
BoehmannY, EnterleinS, RandolfA, MühlbergerE. A reconstituted replication and transcription system for Ebola virus Reston and comparison with Ebola virus Zaire. Virology2005; 332:406–417 [View Article]
TheriaultS, GrosethA, NeumannG, KawaokaY, FeldmannH. Rescue of Ebola virus from cDNA using heterologous support proteins. Virus Res2004; 106:43–50 [View Article]
WendtL, KämperL, SchmidtML, MettenleiterTC, HoenenT. Analysis of a putative late domain using an Ebola virus transcription and replication-competent virus-like particle system. J Infect Dis2018; 218:S355–S359 [View Article]
BornholdtZA, NodaT, AbelsonDM, HalfmannP, WoodMR et al. Structural rearrangement of Ebola virus VP40 begets multiple functions in the virus life cycle. Cell2013; 154:763–774 [View Article]
HoenenT, BiedenkopfN, ZieleckiF, JungS, GrosethA et al. Oligomerization of Ebola virus VP40 is essential for particle morphogenesis and regulation of viral transcription. J Virol2010; 84:7053–7063 [View Article]
FuchsJ, HölzerM, SchillingM, PatzinaC, SchoenA et al. Evolution and antiviral specificities of interferon-induced Mx proteins of bats against Ebola, influenza, and other RNA viruses. J Virol2017; 91: 01 08 2017 [View Article]
McCarthySDS, Majchrzak-KitaB, RacineT, KozlowskiHN, BakerDP et al. A rapid screening assay identifies monotherapy with Interferon-ß and combination therapies with nucleoside analogs as effective inhibitors of Ebola virus. PLoS Negl Trop Dis2016; 10:e0004364 [View Article]
WangZ-Y, GuoZ-D, LiJ-M, ZhaoZ-Z, FuY-Y et al. Genome-wide search for competing endogenous RNAs responsible for the effects induced by Ebola virus replication and transcription using a trVLP system. Front Cell Infect Microbiol2017; 7:479 [View Article]
WangZ, LiJ, FuY, ZhaoZ, ZhangC et al. A rapid screen for host-encoded miRNAs with inhibitory effects against Ebola virus using a transcription- and replication-competent virus-like particle system. Int J Mol Sci2018; 19:E1488 16 May 2018 [View Article]
LeeN, ShumD, KönigA, KimH, HeoJ et al. High-throughput drug screening using the Ebola virus transcription- and replication-competent virus-like particle system. Antiviral Res2018; 158:226–237 [View Article]
VolchkovVE, VolchkovaVA, MuhlbergerE, KolesnikovaLV, WeikM et al. Recovery of infectious Ebola virus from complementary DNA: RNA editing of the GP gene and viral cytotoxicity. Science2001; 291:1965–1969 [View Article]
NeumannG, FeldmannH, WatanabeS, LukashevichI, KawaokaY. Reverse genetics demonstrates that proteolytic processing of the Ebola virus glycoprotein is not essential for replication in cell culture. J Virol2002; 76:406–410 [View Article]
FischerK, JabatyJ, SulukuR, StreckerT, GrosethA et al. Serological evidence for the circulation of Ebolaviruses in pigs from Sierra Leone. J Infect Dis2018; 218:S305-S311 [View Article]
ManhartWA, PachecoJR, HumeAJ, CresseyTN, DeflubéLR et al. A chimeric Lloviu virus minigenome system reveals that the bat-derived filovirus replicates more similarly to ebolaviruses than marburgviruses. Cell Rep2018; 24:2573–2580 [View Article]
MartinS, ChiramelAI, SchmidtML, ChenY-C, WhittN et al. A genome-wide siRNA screen identifies a druggable host pathway essential for the Ebola virus life cycle. Genome Med2018; 10:58 [View Article]
FuC, DonovanWP, Shikapwashya-HasserO, YeX, ColeRH. Hot fusion: an efficient method to clone multiple DNA fragments as well as inverted repeats without ligase. PLoS One2014; 9:e115318 [View Article]
SchmidtML, TewsBA, GrosethA, HoenenT. Generation and optimization of a green fluorescent protein-expressing transcription and replication-competent virus-like particle system for Ebola virus. J Infect Dis2018
SchindelinJ, RuedenCT, HinerMC, EliceiriKW. The ImageJ ecosystem: an open platform for biomedical image analysis. Mol Reprod Dev2015; 82:518–529 [View Article]
TamuraK, NeiM. Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol Biol Evol1993; 10:512–526 [View Article]
TamuraK. Estimation of the number of nucleotide substitutions when there are strong transition-transversion and G+C-content biases. Mol Biol Evol1992; 9:678–687 [View Article]