RESUMEN

Lithified layers of complex microbial mats known as microbialites are ubiquitous in the fossil record, and modern forms are increasingly identified globally. A key challenge to developing an understanding of microbialite formation and environmental role is how to investigate complex and diverse communities in situ. We selected living, layered microbialites (stromatolites) in a peritidal environment near Schoenmakerskop, Eastern Cape, South Africa to conduct a spatial survey mapping the composition and small molecule production of the microbial communities from environmental samples. Substrate core samples were collected from nine sampling stations ranging from the upper point of the freshwater inflow to the lower marine interface where tidal overtopping takes place. Substrate cores provided material for parallel analyses of microbial community diversity by 16S rRNA gene amplicon sequencing and metabolomics using LC-MS2. Species and metabolite diversities were correlated, and prominent specialized metabolites were targeted for preliminary characterization. A new series of cyclic hexadepsipeptides, named ibhayipeptolides, was most abundant in substrate cores of submerged microbialites. These results demonstrate the detection and identification of metabolites from mass-limited environmental samples and contribute knowledge about microbialite chemistry and biology, which facilitates future targeted studies of specialized metabolite function and biosynthesis.

Asunto(s)

Metabolómica , Metabolómica/métodos , Sudáfrica , ARN Ribosómico 16S/genética , Sedimentos Geológicos/microbiología , Depsipéptidos/biosíntesis , Depsipéptidos/química , Bacterias/metabolismo , Bacterias/genética , Bacterias/clasificación

RESUMEN

While anthropogenic pollution is a major threat to aquatic ecosystem health, our knowledge of the presence of xenobiotics in coastal Dissolved Organic Matter (DOM) is still relatively poor. This is especially true for water bodies in the Global South with limited information gained mostly from targeted studies that rely on comparison with authentic standards. In recent years, non-targeted tandem mass spectrometry has emerged as a powerful tool to collectively detect and identify pollutants and biogenic DOM components in the environment, but this approach has yet to be widely utilized for monitoring ecologically important aquatic systems. In this study we compared the DOM composition of Algoa Bay, Eastern Cape, South Africa, and its two estuaries. The Swartkops Estuary is highly urbanized and severely impacted by anthropogenic pollution, while the Sundays Estuary is impacted by commercial agriculture in its catchment. We employed solid-phase extraction followed by liquid chromatography tandem mass spectrometry to annotate more than 200 pharmaceuticals, pesticides, urban xenobiotics, and natural products based on spectral matching. The identification with authentic standards confirmed the presence of methamphetamine, carbamazepine, sulfamethoxazole, N-acetylsulfamethoxazole, imazapyr, caffeine and hexa(methoxymethyl)melamine, and allowed semi-quantitative estimations for annotated xenobiotics. The Swartkops Estuary DOM composition was strongly impacted by features annotated as urban pollutants including pharmaceuticals such as melamines and antiretrovirals. By contrast, the Sundays Estuary exhibited significant enrichment of molecules annotated as agrochemicals widely used in the citrus farming industry, with predicted concentrations for some of them exceeding predicted no-effect concentrations. This study provides new insight into anthropogenic impact on the Algoa Bay system and demonstrates the utility of non-targeted tandem mass spectrometry as a sensitive tool for assessing the health of ecologically important coastal ecosystems and will serve as a valuable foundation for strategizing long-term monitoring efforts.

Asunto(s)

Materia Orgánica Disuelta , Contaminantes Ambientales , Ecosistema , Estuarios , Bahías , Ríos/química , Agricultura , Preparaciones Farmacéuticas

RESUMEN

Pyrroloiminoquinones are a group of cytotoxic alkaloids most commonly isolated from marine sponges. Structurally, they are based on a tricyclic pyrrolo[4,3,2-de]quinoline core and encompass marine natural products such as makaluvamines, tsitsikammamines and discorhabdins. These diverse compounds are known to exhibit a broad spectrum of biological activities including anticancer, antiplasmodial, antimicrobial, antifungal and antiviral activities as well as the inhibition of several key cellular enzymes. The resurgence of interest in pyrroloiminoquinones and the convoluted understanding regarding their biological activities have prompted this review. Herein, we provided a concise summary of key findings and recent developments pertaining to their structural diversity, distribution, biogenesis, and their potential as chemical probes for drug development, including a discussion of promising synthetic analogs.

Asunto(s)

Alcaloides , Antineoplásicos , Productos Biológicos , Poríferos , Pirroliminoquinonas , Animales , Pirroliminoquinonas/química , Pirroliminoquinonas/farmacología , Poríferos/química , Antineoplásicos/química , Alcaloides/química , Descubrimiento de Drogas

RESUMEN

The fossil record indicates that the earliest evidence of extant marine sponges (phylum Porifera) existed during the Cambrian explosion and that their symbiosis with microbes may have begun in their extinct ancestors during the Precambrian period. Many symbionts have adapted to their sponge host, where they perform specific, specialized functions. There are also widely distributed bacterial taxa such as Poribacteria, SAUL, and Tethybacterales that are found in a broad range of invertebrate hosts. Here, we added 11 new genomes to the Tethybacterales order, identified a novel family, and show that functional potential differs between the three Tethybacterales families. We compare the Tethybacterales with the well-characterized Entoporibacteria and show that these symbionts appear to preferentially associate with low-microbial abundance (LMA) and high-microbial abundance (HMA) sponges, respectively. Within these sponges, we show that these symbionts likely perform distinct functions and may have undergone multiple association events, rather than a single association event followed by coevolution. IMPORTANCE Marine sponges often form symbiotic relationships with bacteria that fulfil a specific need within the sponge holobiont, and these symbionts are often conserved within a narrow range of related taxa. To date, there exist only three known bacterial taxa (Entoporibacteria, SAUL, and Tethybacterales) that are globally distributed and found in a broad range of sponge hosts, and little is known about the latter two. We show that the functional potential of broad-host range symbionts is conserved at a family level and that these symbionts have been acquired several times over evolutionary history. Finally, it appears that the Entoporibacteria are associated primarily with high-microbial abundance sponges, while the Tethybacterales associate with low-microbial abundance sponges.

Asunto(s)

Bacterias/genética , Genómica , Especificidad del Huésped , Poríferos/microbiología , Simbiosis , Animales , Bacterias/clasificación , Microbiota , Filogenia , ARN Ribosómico 16S , Agua de Mar/microbiología

RESUMEN

Correction for 'Antiviral drug discovery: preparing for the next pandemic' by Catherine S. Adamson et al., Chem. Soc. Rev., 2021, 50, 3647-3655, DOI: .

RESUMEN

Clinically approved antiviral drugs are currently available for only 10 of the more than 220 viruses known to infect humans. The SARS-CoV-2 outbreak has exposed the critical need for compounds that can be rapidly mobilised for the treatment of re-emerging or emerging viral diseases, while vaccine development is underway. We review the current status of antiviral therapies focusing on RNA viruses, highlighting strategies for antiviral drug discovery and discuss the challenges, solutions and options to accelerate drug discovery efforts.

Asunto(s)

Antivirales/farmacología , Tratamiento Farmacológico de COVID-19 , Descubrimiento de Drogas/métodos , Terapia Molecular Dirigida/métodos , Pandemias/prevención & control , ARN Viral/antagonistas & inhibidores , Antivirales/química , Productos Biológicos/química , Productos Biológicos/farmacología , COVID-19/prevención & control , COVID-19/virología , Inhibidores de Proteasa de Coronavirus/química , Inhibidores de Proteasa de Coronavirus/farmacología , Humanos , Simulación del Acoplamiento Molecular , Inhibidores de la Síntesis del Ácido Nucleico/química , Inhibidores de la Síntesis del Ácido Nucleico/farmacología , ARN Viral/química , ARN Viral/genética , ARN Viral/metabolismo , SARS-CoV-2/química , SARS-CoV-2/efectos de los fármacos , SARS-CoV-2/enzimología , SARS-CoV-2/genética , Bibliotecas de Moléculas Pequeñas/química , Bibliotecas de Moléculas Pequeñas/farmacología

RESUMEN

Sponges of the Latrunculiidae family produce bioactive pyrroloiminoquinone alkaloids including makaluvamines, discorhabdins, and tsitsikammamines. The aim of this study was to use LC-ESI-MS/MS-driven molecular networking to characterize the pyrroloiminoquinone secondary metabolites produced by six latrunculid species. These are Tsitsikamma favus, Tsitsikamma pedunculata, Cyclacanthia bellae, and Latrunculia apicalis as well as the recently discovered species, Tsitsikamma nguni and Tsitsikamma michaeli. Organic extracts of 43 sponges were analyzed, revealing distinct species-specific chemical profiles. More than 200 known and unknown putative pyrroloiminoquinones and related compounds were detected, including unprecedented makaluvamine-discorhabdin adducts and hydroxylated discorhabdin I derivatives. The chemical profiles of the new species T. nguni closely resembled those of the known T. favus (chemotype I), but with a higher abundance of tsitsikammamines vs. discorhabdins. T. michaeli sponges displayed two distinct chemical profiles, either producing mostly the same discorhabdins as T. favus (chemotype I) or non- or monobrominated, hydroxylated discorhabdins. C. bellae and L. apicalis produced similar pyrroloiminoquinone chemistry to one another, characterized by sulfur-containing discorhabdins and related adducts and oligomers. This study highlights the variability of pyrroloiminoquinone production by latrunculid species, identifies novel isolation targets, and offers fundamental insights into the collision-induced dissociation of pyrroloiminoquinones.

Asunto(s)

Biodiversidad , Redes Reguladoras de Genes/fisiología , Poríferos/genética , Pirroliminoquinonas/aislamiento & purificación , Animales

RESUMEN

In the geological record, fossil phosphatic stromatolites date back to the Great Oxidation Event in the Paleoproterozoic, but living phosphatic stromatolites have not been described previously. Here, we report on cyanobacterial stromatolites in a supratidal freshwater environment at Cape Recife, South African southern coast, precipitating Ca carbonate alternating with episodes of Ca phosphate deposition. In their structure and composition, the living stromatolites from Cape Recife closely resemble their fossilized analogues, showing phosphatic zonation, microbial casts, tunnel structures and phosphatic crusts of biogenic origin. The microbial communities appear to be also similar to those proposed to have formed fossil phosphatic stromatolites. Phosphatic domains in the material from Cape Recife are spatially and texturally associated with carbonate precipitates, but form distinct entities separated by sharp boundaries. Electron Probe Micro-Analysis shows that Ca/P ratios and the overall chemical compositions of phosphatic precipitates are in the range of octacalcium phosphate, amorphous tricalcium phosphate and apatite. The coincidence in time of the emergence of phosphatic stromatolites in the fossil record with a major episode of atmospheric oxidation led to the assumption that at times of increased oxygen release the underlying increased biological production may have been linked to elevated phosphorus availability. The stromatolites at Cape Recife, however, form in an environment where ambient phosphorus concentrations do not exceed 0.28 µM, one to two orders of magnitude below the previously predicted minimum threshold of >5 µM for biogenic phosphate precipitation in paleo-systems. Accordingly, we contest the previously proposed suitability of phosphatic stromatolites as a proxy for high ambient phosphate concentrations in supratidal to shallow ocean settings in earth history.

Asunto(s)

Cianobacterias , Fósiles , Fósforo , Sedimentos Geológicos , Geología , Fosfatos , Fósforo/análisis

RESUMEN

Stromatolites are complex microbial mats that form lithified layers. Fossilized stromatolites are the oldest evidence of cellular life on Earth, dating back over 3.4 billion years. Modern stromatolites are relatively rare but may provide clues about the function and evolution of their ancient counterparts. In this study, we focus on peritidal stromatolites occurring at Cape Recife and Schoenmakerskop on the southeastern South African coastline, the former being morphologically and structurally similar to fossilized phosphatic stromatolites formations. Using assembled shotgun metagenomic analysis, we obtained 183 genomic bins, of which the most dominant taxa were from the Cyanobacteria phylum. We identified functional gene sets in genomic bins conserved across two geographically isolated stromatolite formations, which included relatively high copy numbers of genes involved in the reduction of nitrates and phosphatic compounds. Additionally, we found little evidence of Archaeal species in these stromatolites, suggesting that they may not play an important role in peritidal stromatolite formations, as proposed for hypersaline formations.

Asunto(s)

Cianobacterias , Sedimentos Geológicos , Archaea , Cianobacterias/genética , Genoma Bacteriano , Sedimentos Geológicos/microbiología , Metagenómica

RESUMEN

Tetraviruses are a group of relatively unknown small RNA viruses with particles that display a characteristic T=4 capsid architecture. Tetraviruses are classified into three families, the Alphatetraviridae, Permutotetraviridae and Carmotetraviridae, according to the divergent characteristics of their respective viral replicases. Tetraviruses generally infect the larvae of lepidopteran insect species, many of which are important agricultural pests and, until recently, were thought to have an unusually narrow host range and tissue tropism. The development of experimental systems for studying the viral infectious life cycle in tissue culture has permitted the extension of the virus host range to mammalian cells and plants. This chapter will review recent advances in the understanding of the biology of tetraviruses, highlighting new information on the expression and functional characterisation of viral proteins and the development of biological systems for elucidating the molecular mechanisms of infection, viral replication and host range.

Asunto(s)

Especificidad del Huésped , Estadios del Ciclo de Vida , Infecciones por Virus ARN/virología , Virus ARN/fisiología , Tropismo Viral , Cápside/metabolismo , Código de Barras del ADN Taxonómico , Regulación Viral de la Expresión Génica , Genoma Viral , Genómica/métodos , Virus ARN/clasificación , Ensamble de Virus , Acoplamiento Viral , Internalización del Virus , Replicación Viral

RESUMEN

The genus Tsitsikamma Samaai & Kelly, 2002 is to date exclusively reported from South Africa. Three species are known from the southern coast: Tsitsikamma favus Samaai & Kelly, 2002, from the Garden Route National Park Tsitsikamma Marine Protected Area (MPA) and Algoa Bay; T. pedunculata Samaai, Gibbons, Kelly and Davies-Coleman, 2003, collected from Cape Recife in St. Francis Bay, and T. scurra Samaai, Gibbons, Kelly and Davies-Coleman, 2003, collected from a wreck site in a small bay west of Hout Bay on the west coast of South Africa. Here two new species are described: Tsitsikamma michaeli Parker-Nance, sp. nov., a small green purse-like species, collected from Algoa Bay, and Tsitsikamma nguni Parker-Nance, sp. nov., from The Garden Route National Park, Tsitsikamma MPA. Additional morphological characteristics, spicule morphology, and distribution records are provided for T. favus and T. pedunculata from Algoa Bay. The phylogenetic relationship of these five Tsitsikamma species is investigated.

RESUMEN

INTRODUCTION: Emerging viral diseases, most of which are zoonotic, pose a significant threat to global health. There is a critical need to identify potential new viral pathogens and the challenge is to identify the reservoirs from which these viruses might emerge. Deep sequencing of invertebrate transcriptomes has revealed a plethora of viruses, many of which represent novel lineages representing both plant and animal viruses and little is known about the potential threat that these viruses pose. METHODS: Providence virus, an insect virus, was used to establish a productive infection in Vigna unguiculata (cowpea) plants. Providence virus particles purified from these cowpea plants were used to infect two mammalian cell lines. FINDINGS: Here, we present evidence that Providence virus, a non-enveloped insect RNA virus, isolated from a lepidopteran midgut cell line can establish a productive infection in plants as well as in animal cells. The observation that Providence virus can readily infect both plants and mammalian cell culture lines demonstrates the ability of an insect RNA virus to establish productive infections across two kingdoms, in plants and invertebrate and vertebrate animal cell lines. CONCLUSIONS: The study highlights the potential of phytophagous insects as reservoirs for viral re-assortment and that plants should be considered as reservoirs for emerging viruses that may be potentially pathogenic to humans.

Asunto(s)

Lepidópteros/virología , Células Vegetales/virología , Infecciones por Virus ARN/metabolismo , Vigna/virología , Animales , Células HeLa , Humanos , Células MCF-7 , Virus ARN

RESUMEN

The temperate marine sponge, Tsitsikamma favus, produces pyrroloiminoquinone alkaloids with potential as anticancer drug leads. We profiled the secondary metabolite reservoir of T. favus sponges using HR-ESI-LC-MS/MS-based molecular networking analysis followed by preparative purification efforts to map the diversity of new and known pyrroloiminoquinones and related compounds in extracts of seven specimens. Molecular taxonomic identification confirmed all sponges as T. favus and five specimens (chemotype I) were found to produce mainly discorhabdins and tsitsikammamines. Remarkably, however, two specimens (chemotype II) exhibited distinct morphological and chemical characteristics: the absence of discorhabdins, only trace levels of tsitsikammamines and, instead, an abundance of unbranched and halogenated makaluvamines. Targeted chromatographic isolation provided the new makaluvamine Q, the known makaluvamines A and I, tsitsikammamine B, 14-bromo-7,8-dehydro-3-dihydro-discorhabdin C, and the related pyrrolo-ortho-quinones makaluvamine O and makaluvone. Purified compounds displayed different activity profiles in assays for topoisomerase I inhibition, DNA intercalation and antimetabolic activity against human cell lines. This is the first report of makaluvamines from a Tsitsikamma sponge species, and the first description of distinct chemotypes within a species of the Latrunculiidae family. This study sheds new light on the putative pyrroloiminoquinone biosynthetic pathway of latrunculid sponges.

Asunto(s)

Poríferos/metabolismo , Pirroliminoquinonas/química , Animales , Antimetabolitos Antineoplásicos/química , Antimetabolitos Antineoplásicos/aislamiento & purificación , Antimetabolitos Antineoplásicos/farmacología , Vías Biosintéticas , Supervivencia Celular/efectos de los fármacos , Cromatografía Líquida de Alta Presión/métodos , ADN/química , ADN/efectos de los fármacos , ADN-Topoisomerasas de Tipo I/metabolismo , Pruebas de Enzimas , Células HEK293 , Células HeLa , Humanos , Sustancias Intercalantes/química , Sustancias Intercalantes/aislamiento & purificación , Sustancias Intercalantes/farmacología , Estructura Molecular , Pirroliminoquinonas/aislamiento & purificación , Pirroliminoquinonas/metabolismo , Pirroliminoquinonas/farmacología , Espectrometría de Masas en Tándem/métodos , Inhibidores de Topoisomerasa I/química , Inhibidores de Topoisomerasa I/aislamiento & purificación , Inhibidores de Topoisomerasa I/metabolismo , Inhibidores de Topoisomerasa I/farmacología

RESUMEN

Mesoscale variability and associated eddy fluxes play crucial roles in ocean circulation dynamics and the ecology of the upper ocean. In doing so, these features are biologically important, providing a mechanism for the mixing and exchange of nutrients and biota within the ocean. Transient mesoscale eddies in the Southern Ocean are known to relocate zooplankton communities across the Antarctic Circumpolar Current (ACC) and are important foraging grounds for marine top predators. In this study we investigated the role of cyclonic and anti-cyclonic eddies formed at the South-West Indian Ridge on the spatial variability and diversity of microbial communities. We focused on two contrasting adjacent eddies within the Antarctic Polar Frontal Zone to determine how these features may influence the microbial communities within this region. The water masses and microbiota of the two eddies, representative of a cyclonic cold core from the Antarctic zone and an anti-cyclonic warm-core from the Subantarctic zone, were compared. The data reveal that the two eddies entrain distinct microbial communities from their points of origin that are maintained for up to ten months. Our findings highlight the ecological impact that changes, brought by the translocation of eddies across the ACC, have on microbial diversity.

Asunto(s)

Ecosistema , Océanos y Mares , Microbiología del Agua , Movimientos del Agua , Regiones Antárticas , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , ARN Ribosómico 16S/genética

RESUMEN

Sponges are important sources of bioactive secondary metabolites. These compounds are frequently synthesized by bacterial symbionts, which may be recruited from the surrounding seawater or transferred to the sponge progeny by the parent. In this study, we investigated the bacterial communities associated with the sponge Tethya rubra Samaai and Gibbons 2005. Sponge specimens were collected from Evans Peak and RIY Banks reefs in Algoa Bay, South Africa and taxonomically identified by spicule analysis and molecular barcoding. Crude chemical extracts generated from individual sponges were profiled by ultraviolet high performance liquid chromatography (UV-HPLC) and subjected to bioactivity assays in mammalian cells. Next-generation sequencing analysis of 16S rRNA gene sequences was used to characterize sponge-associated bacterial communities. T. rubra sponges collected from the two locations were morphologically and genetically indistinguishable. Chemical extracts from sponges collected at RIY banks showed mild inhibition of the metabolic activity of mammalian cells and their UV-HPLC profiles were distinct from those of sponges collected at Evans Peak. Similarly, the bacterial communities associated with sponges from the two locations were distinct with evidence of vertical transmission of symbionts from the sponge parent to its embryos. We conclude that these distinct bacterial communities may be responsible for the differences observed in the chemical profiles of the two Algoa Bay T. rubra Samaai and Gibbons 2005 populations.

Asunto(s)

Bacterias/genética , Bahías/microbiología , Poríferos/microbiología , Animales , Biodiversidad , ADN Bacteriano/genética , Filogenia , ARN Ribosómico 16S/genética , Agua de Mar/microbiología , Análisis de Secuencia de ADN/métodos , Sudáfrica

RESUMEN

The Latrunculiidae are a family of cold water sponges known for their production of bioactive pyrroloiminoquinone alkaloids. Previously it was shown that the bacterial community associated with a Tsitsikamma sponge species comprises unusual bacterial taxa and is dominated by a novel Betaproteobacterium. Here, we have characterized the bacterial communities associated with six latrunculid species representing three genera (Tsitsikamma, Cyclacanthia, and Latrunculia) as well as a Mycale species, collected from Algoa Bay on the South African southeast coast. The bacterial communities of all seven sponge species were dominated by a single Betaproteobacterium operational taxonomic unit (OTU0.03 ), while a second OTU0.03 was dominant in the Mycale sp. The Betaproteobacteria OTUs from the different latrunculid sponges are closely related and their phylogenetic relationship follows that of their hosts. We propose that the latrunculid Betaproteobacteria OTUs are members of a specialized group of sponge symbionts that may have coevolved with their hosts. A single dominant Spirochaetae OTU0.03 was present in the Tsitsikamma and Cyclacanthia sponge species, but absent from the Latrunculia and Mycale sponges. This study sheds new light on the interactions between latrunculid sponges and their bacterial communities and may point to the potential involvement of dominant symbionts in the biosynthesis of the bioactive secondary metabolites.

Asunto(s)

Betaproteobacteria/aislamiento & purificación , ADN Bacteriano/genética , Microbiota/genética , Poríferos/clasificación , Poríferos/microbiología , Animales , Secuencia de Bases , Betaproteobacteria/clasificación , Betaproteobacteria/genética , Amplificación de Genes , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Sudáfrica , Simbiosis

RESUMEN

Tetraviruses are small, non-enveloped, RNA viruses that exclusively infect lepidopteran insects. Their particles comprise 240 copies of a single capsid protein precursor (CP), which undergoes autoproteolytic cleavage during maturation. The molecular mechanisms of capsid assembly and maturation are well understood, but little is known about the viral infectious lifecycle due to a lack of tissue culture cell lines that are susceptible to tetravirus infection. We show here that binding and entry of the alphatetravirus, Helicoverpa armigera stunt virus (HaSV), is triggered by alkaline pH. At pH 9.0, wild-type HaSV virus particles undergo conformational changes that induce membrane-lytic activity and binding to Spodoptera frugiperda Sf9 cells. Binding is followed by entry and infection, with virus replication complexes detected by immunofluorescence microscopy within 2h post-infection and the CP after 12h. HaSV particles produced in S. frugiperda Sf9 cells are infectious. Helicoverpa armigera larval virus biofeed assays showed that pre-treatment with the V-ATPase inhibitor, Bafilomycin A1, resulted in a 50% decrease in larval mortality and stunting, while incubation of virus particles at pH 9.0 prior to infection restored infectivity. Together, these data show that HaSV, and likely other tetraviruses, requires the alkaline environment of the lepidopteran larval midgut for binding and entry into host cells.

Asunto(s)

Concentración de Iones de Hidrógeno , Virus de Insectos/fisiología , Virus ARN/fisiología , Acoplamiento Viral , Internalización del Virus , Animales , Cápside/química , Cápside/metabolismo , Proteínas de la Cápside/química , Proteínas de la Cápside/metabolismo , Línea Celular , Virus de Insectos/ultraestructura , Modelos Moleculares , Conformación Proteica , Virus ARN/ultraestructura , Células Sf9 , Spodoptera/virología , Replicación Viral

RESUMEN

Tetraviruses are small, positive (+ve)-sense ssRNA viruses that infect the midgut cells of lepidopteran larvae. Providence virus (PrV) is the only member of the family Carmotetraviridae (previously Tetraviridae). PrV particles exhibit the characteristic tetraviral T=4 icosahedral symmetry, but PrV is distinct from other tetraviruses with respect to genome organization and viral non-structural proteins. Currently, PrV is the only tetravirus known to infect and replicate in lepidopteran cell culture lines. In this report we demonstrate, using immunofluorescence microscopy, that PrV infects and replicates in a human tissue culture cell line (HeLa), producing infectious virus particles. We also provide evidence for PrV replication in vitro in insect, mammalian and plant cell-free systems. This study challenges the long-held view that tetraviruses have a narrow host range confined to one or a few lepidopteran species and highlights the need to consider the potential for apparently non-infectious viruses to be transferred to new hosts in the laboratory.

Asunto(s)

Virus de Insectos/fisiología , Virus ARN/fisiología , Animales , Línea Celular , Especificidad del Huésped , Humanos , Virus de Insectos/genética , Virus de Insectos/aislamiento & purificación , Insectos/virología , Mamíferos/virología , Virus ARN/genética , Virus ARN/aislamiento & purificación , Replicación Viral

RESUMEN

Alphatetraviruses are small (+) ssRNA viruses with non-enveloped, icosahedral, T=4 particles that assemble from 240 copies of a single capsid protein precursor. This study is focused on the mechanisms underlying selection and packaging of genomic vRNAs by Helicoverpa armigera stunt virus. We demonstrate that the viral protein, p17, is packaged at low levels (between 4 and 8 copies per capsid) raising the possibility of icosahedral asymmetry in wild-type particles. p17 promotes packaging of vRNA2 by virus-like particles (VLPs) generated from plasmid-expressed vRNA2. The 5' and 3' UTRs of RNA2 are not required for encapsidation. VLPs produced by recombinant baculoviruses package vRNA2 at detectable levels even in the absence of p17 and apparently excluding baculoviral transcripts. This suggests a role for p17 in vRNA selectivity. This is one of few examples of the packaging of a minor non-structural protein by (+) ssRNA animal viruses.

Asunto(s)

Virus de Insectos/fisiología , Lepidópteros/virología , Virus ARN/fisiología , ARN Viral/metabolismo , Ensamble de Virus , Animales , Cápside/metabolismo , Virus de Insectos/genética , Virus ARN/genética , ARN Viral/genética , Proteínas Virales/genética , Proteínas Virales/metabolismo

Asunto(s)

Peces/genética , Genoma , Animales , Historia del Siglo XX , Historia del Siglo XXI