Streptovaricins inhibit Focus Formation by MSV (MLV) Complex

We recently reported that the streptovaricins inhibit the reaction by which DNA is transcribed from the RNA template resident in murine leukaemia virions (MLV)¹. The reports^{2, 3} which first indicated that this DNA polymerase is present in oncogenic RNA viruses have been confirmed and extended^4–8. The enzyme provides a mechanism whereby an RNA virus can insert stable genetic information into a host chromosome. Gallo and co-workers described an RNA dependent DNA polymerase in lymphoblastic leukaemic cells which was inhibited by N-demethylrifampicin⁹ and this antibiotic, together with a number of other rifamycin derivatives, also inhibited the oncogenic viral DNA polymerase¹⁰. Like the streptovaricins, the rifamycins are ansa macrolide antibiotics (Fig. 1).     

Rat C-Type Virus induced in Rat Sarcoma Cells by 5-Bromodeoxyuridine

HALOGENATED derivatives of uridine are highly effective inducers of latent C-type RNA viruses^1,2 and have been successfully used to induce viruses identical to, or similar to, the C-type RNA tumour viruses in mouse, rat and human cells^3–6. In previous experiments we used 5-bromodeoxyuridine (BrUdR) for induction of focus-forming virus in non-productive rat cells that have been transformed by mouse sarcoma virus². We describe here the induction of a C-type RNA virus in the cells of the rat tumour cell line XC, which contains the Rous sarcoma virus genome⁷. The induced virus possesses the group specific (gs) antigens of rat C-type viruses but not those of chicken C-type viruses.     

Lymphocytic Choriomeningitis Virus RNAs

IT has been proposed that lymphocytic choriomeningitis (LCM) virus and immunologically related viruses should be placed in a new taxonomic group, with LCM virus as the prototype virus^1,2. They were called arenoviruses because they contain, as a unique feature, electron-dense, sand-like or ribo-some-like granules². The LCM virus contains RNA and we have previously reported that this RNA can be separated into three components by density gradient centrifugation³.     

Large-scale isolation and partial purification of type C RNA viruses on hydroxyapatite. 1. Biochemical characterization.

A chromatographic procedure utilizing common laboratory equipment and based on the batchwise adsorption of type C RNA virus onto hydroxyapatite for the concentration and partial purification of viruses from large volumes of tissue culture fluid has been developed. This procedure provides an alternative to the use of elaborate and expensive high-speed zonal ultracentrifuge equipment. The viruses obtained by this procedure have a buoyant density of 1.16 g/cm³, contain 70 S RNA, an RNA-directed DNA polymerase (reverse transeriptase), surface glycoproteins ($\text{GP}\text{69}\text{71}$), and the internal viral specific polypeptides p10 to 15 and p27 or p30.     

Inhibitor of the DNA-dependent DNA Polymerase of Some RNA Tumour Viruses in Feline Sera

AN inhibitor of the RNA-dependent DNA polymerases^1,2 of mammalian C-type viruses was found in sera from rats bearing transplantable tumours, induced by murine C-type RNA tumour viruses^3,4. Partially purified polymerases of murine leukaemia virus³ and feline leukaemia virus (FeLV)⁴ were shown to be antigenic in rabbits and a rat, respectively. We have detected an inhibitor of the DNA-dependent DNA polymerase^5,6 of feline and murine C-type viruses in the sera of cats inoculated in utero and/or postnatally with the Gardner-Arnstein strain of feline sarcoma virus (FSV)⁷ and in the sera of cats bearing spontaneous sarcomas, lymphomas or carcinomas.     

Expression of a New Complement-fixing Antigen reactive with Murine Sarcoma Virus Rat Antiserum in Rat Cells transformed by Polyoma Virus

WE reported accelerated transformation by DNA viruses (SV40 and polyoma) in rat embryo (RE) cells chronically infected with a C-type RNA virus^1,2. Recently we found in RE cells transformed by polyoma virus a new complement-fixing (CF) antigen detectable by rat antisera having broad reactivity with the various intraspecies and interspecies antigens of the RNA tumour viruses^3–8; this antigen, however, was distinct from the murine intraspecies and interspecies group-specific (gs) antigens both immunologically and by virtue of other properties. It is also distinct from the polyoma virion (capsid) and tumour (“T”) antigens.     

RNA Dependent DNA Polymerase in C Type Particles from Normal Rat Thymus Cultures

THE isolation and characterization of C type particles released from normal rat thymus cell cultures has been described^{1, 2}. The detection of RNA dependent DNA polymerase activity in these C type particles is reported here. Some properties of the enzymatic activities found in C type particles isolated from normal rat thymus cultures are described and compared with DNA polymerase found in isolated Moloney leukaemia virus (MLV) particles purified from the growth media of rat thymus cultures infected chronically with MLV². The sensitivity to actinomycin D is different in the two kinds of C type particles, suggesting that the DNA polymerases are not identical.     

Analysis of the Solvent Accessibility of Cysteine Residues on Maize rayado fino virus Virus-like Particles Produced in Nicotiana benthamiana Plants and Cross-linking of Peptides to VLPs

Mimicking and exploiting virus properties and physicochemical and physical characteristics holds promise to provide solutions to some of the world''s most pressing challenges. The sheer range and types of viruses coupled with their intriguing properties potentially give endless opportunities for applications in virus-based technologies. Viruses have the ability to self- assemble into particles with discrete shape and size, specificity of symmetry, polyvalence, and stable properties under a wide range of temperature and pH conditions. Not surprisingly, with such a remarkable range of properties, viruses are proposed for use in biomaterials ⁹, vaccines ^{14, 15}, electronic materials, chemical tools, and molecular electronic containers^{4, 5, 10, 11, 16, 18, 12}.In order to utilize viruses in nanotechnology, they must be modified from their natural forms to impart new functions. This challenging process can be performed through several mechanisms including genetic modification of the viral genome and chemically attaching foreign or desired molecules to the virus particle reactive groups ⁸. The ability to modify a virus primarily depends upon the physiochemical and physical properties of the virus. In addition, the genetic or physiochemical modifications need to be performed without adversely affecting the virus native structure and virus function. Maize rayado fino virus (MRFV) coat proteins self-assemble in Escherichia coli producing stable and empty VLPs that are stabilized by protein-protein interactions and that can be used in virus-based technologies applications ⁸. VLPs produced in tobacco plants were examined as a scaffold on which a variety of peptides can be covalently displayed ¹³. Here, we describe the steps to 1) determine which of the solvent-accessible cysteines in a virus capsid are available for modification, and 2) bioconjugate peptides to the modified capsids. By using native or mutationally-inserted amino acid residues and standard coupling technologies, a wide variety of materials have been displayed on the surface of plant viruses such as, Brome mosaic virus ³, Carnation mottle virus ¹², Cowpea chlorotic mottle virus ⁶, Tobacco mosaic virus ¹⁷, Turnip yellow mosaic virus ¹, and MRFV ¹³.     

Whitefly‐transmitted RNA viruses that affect intensive vegetable production

Intensive vegetable production is constantly facing the emergence of new viral diseases. Apart from the intrinsic features of viruses as plant pathogens, the highly dynamic turnover of cultivars and cultural practices, and the global trade of seeds and products characteristic of intensive vegetable production may favour the emergence of new viruses, as well as the expansion of the geographical range of vectors responsible for their dissemination. Indeed, the efficient transmission of viruses plays a major role in the impact and outcome of viral epidemics. Whiteflies (Hemiptera: Aleyrodidae) that belong to the genera Bemisia and Trialeurodes are efficient virus vectors. Whiteflies transmit viruses of at least four genera, one of DNA viruses, the genus Begomovirus, and three of RNA viruses, Crinivirus, Ipomovirus and Torradovirus. Begomoviruses have been the subject of recent reviews. In this article we review the genome structure, epidemiology and control of whitefly‐transmitted RNA viruses that belong to the genera Crinivirus, Ipomovirus and Torradovirus, with an extended discussion on the particular viruses within these genera that are currently causing important outbreaks, such are Cucumber vein yellowing virus (CVYV), Cucurbit yellow stunting disorder virus (CYSDV), Tomato chlorosis virus (ToCV) and Tomato torrado virus (ToTV).     

Upper-limit mutation rate estimation for a plant RNA virus

It is generally accepted that mutation rates of RNA viruses are inherently high due to the lack of proofreading mechanisms. However, direct estimates of mutation rate are surprisingly scarce, in particular for plant viruses. Here, based on the analysis of in vivo mutation frequencies in tobacco etch virus, we calculate an upper-bound mutation rate estimation of 3×10⁻⁵ per site and per round of replication; a value which turns out to be undistinguishable from the methodological error. Nonetheless, the value is barely on the lower side of the range accepted for RNA viruses, although in good agreement with the only direct estimate obtained for other plant viruses. These observations suggest that, perhaps, differences in the selective pressures operating during plant virus evolution may have driven their mutation rates towards values lower than those characteristic of other RNA viruses infecting bacteria or animals.     

Spontaneous Release of Endogenous Ecotropic Type C Virus from Rat Embryo Cultures

Type C viruses were isolated from embryo cultures of two different rat strains, Sprague-Dawley and Fischer. Both viruses (termed rat leukemia virus, RaLV) were released spontaneously from rat embryo cells, have a density of 1.14 to 1.15 g/cm(3) based on equilibrium sedimentation in sucrose gradients, contain 60-70S RNA, RNA-directed DNA polymerase, and rat type C virus-specific 30,000 molecular-weight-protein determinants. Molecular hybridization studies using the Sprague-Dawley RaLV 60-70S RNA show that the virus-specific nucleotide sequences are present in the DNA of rat embryos. Both Sprague-Dawley and Fischer RaLV can rescue the murine sarcoma virus genome from Kirsten murine sarcoma virus-transformed nonproducer cells and are neutralized by antisera to the RPL strain of RaLV. In contrast to previous RaLV's, these viruses propagate in their own cells of origin as well as in cells of heterologous rat strains.     

A Novel Mycovirus That Is Related to the Human Pathogen Hepatitis E Virus and Rubi-Like Viruses

Previously, we reported that three double-stranded RNA (dsRNA) segments, designated L-, M-, and S-dsRNAs, were detected in Sclerotinia sclerotiorum strain Ep-1PN. Of these, the M-dsRNA segment was derived from the genomic RNA of a potexvirus-like positive-strand RNA virus, Sclerotinia sclerotiorum debilitation-associated RNA virus. Here, we present the complete nucleotide sequence of the L-dsRNA, which is 6,043 nucleotides in length, excluding the poly(A) tail. Sequence analysis revealed the presence of a single open reading frame (nucleotide positions 42 to 5936) that encodes a protein with significant similarity to the replicases of the “alphavirus-like” supergroup of positive-strand RNA viruses. A sequence comparison of the L-dsRNA-encoded putative replicase protein containing conserved methyltransferase, helicase, and RNA-dependent RNA polymerase motifs showed that it has significant sequence similarity to the replicase of Hepatitis E virus, a virus infecting humans. Furthermore, we present convincing evidence that the virus-like L-dsRNA could replicate independently with only a slight impact on growth and virulence of its host. Our results suggest that the L-dsRNA from strain Ep-1PN is derived from the genomic RNA of a positive-strand RNA virus, which we named Sclerotinia sclerotiorum RNA virus L (SsRV-L). As far as we know, this is the first report of a positive-strand RNA mycovirus that is related to a human virus. Phylogenetic and sequence analyses of the conserved motifs of the RNA replicase of SsRV-L showed that it clustered with the rubi-like viruses and that it is related to the plant clostero-, beny- and tobamoviruses and to the insect omegatetraviruses. Considering the fact that these related alphavirus-like positive-strand RNA viruses infect a wide variety of organisms, these findings suggest that the ancestral positive-strand RNA viruses might be of ancient origin and/or they might have radiated horizontally among vertebrates, insects, plants, and fungi.     

Inactivation of DNA Polymerases of Murine Leukaemia Viruses by Calcium Elenolate

THE monoterpene elenolic acid can be isolated, after mild acid hydrolysis, from aqueous extracts of the olive plant (Olea europa)^1,2. The hydrolysate yields crystalline calcium elenolate [(C₁₁H₁₃O₆)₂Ca] which is virucidal in vitro for a number of both DNA and RNA viruses³. It also reduces virus yields from hamsters infected with parainfluenza 3 virus⁴ and has minimal toxicity when administered to animals⁵. I report the inhibition of RNA-dependent DNA polymerases of murine leukaemia viruses by calcium elenolate.     

The Global Trade in Fresh Produce and the Vagility of Plant Viruses: A Case Study in Garlic

As cuisine becomes globalized, large volumes of fresh produce are traded internationally. The potential exists for pathogens infecting fresh produce to hitchhike to new locations and perhaps to establish there. It is difficult to identify them using traditional methods if pathogens are novel, scarce, and/or unexpected. In an attempt to overcome this limitation, we used high-throughput sequencing technology as a means of detecting all RNA viruses infecting garlic (Allium sativum L.) bulbs imported into Australia from China, the USA, Mexico, Argentina and Spain, and those growing in Australia. Bulbs tested were grown over multiple vegetative generations and all were stably infected with one or more viruses, including two species not previously recorded in Australia. Present in various combinations from 10 garlic bulbs were 41 virus isolates representing potyviruses (Onion yellow dwarf virus, Leek yellow stripe virus), carlaviruses (Shallot latent virus, Garlic common latent virus) and allexiviruses (Garlic virus A, B, C, D, and X), for which 19 complete and 22 partial genome sequences were obtained, including the first complete genome sequences of two isolates of GarVD. The most genetically distinct isolates of GarVA and GarVX described so far were identified from Mexico and Argentina, and possible scenarios explaining this are presented. The complete genome sequence of an isolate of the potexvirus Asparagus virus 3 (AV3) was obtained in Australia from wild garlic (A. vineale L.), a naturalized weed. This is first time AV3 has been identified from wild garlic and the first time it has been identified beyond China and Japan. The need for routine generic diagnosis and appropriate legislation to address the risks to primary production and wild plant communities from pathogens spread through the international trade in fresh produce is discussed.     

Genetic analysis of resistance to six virus diseases in a multiple virus-resistant maize inbred line

Key message

Novel and previously known resistance loci for six phylogenetically diverse viruses were tightly clustered on chromosomes 2, 3, 6 and 10 in the multiply virus-resistant maize inbred line, Oh1VI.

Abstract

Virus diseases in maize can cause severe yield reductions that threaten crop production and food supplies in some regions of the world. Genetic resistance to different viruses has been characterized in maize populations in diverse environments using different screening techniques, and resistance loci have been mapped to all maize chromosomes. The maize inbred line, Oh1VI, is resistant to at least ten viruses, including viruses in five different families. To determine the genes and inheritance mechanisms responsible for the multiple virus resistance in this line, F₁ hybrids, F₂ progeny and a recombinant inbred line (RIL) population derived from a cross of Oh1VI and the virus-susceptible inbred line Oh28 were evaluated. Progeny were screened for their responses to Maize dwarf mosaic virus, Sugarcane mosaic virus, Wheat streak mosaic virus, Maize chlorotic dwarf virus, Maize fine streak virus, and Maize mosaic virus. Depending on the virus, dominant, recessive, or additive gene effects were responsible for the resistance observed in F₁ plants. One to three gene models explained the observed segregation of resistance in the F₂ generation for all six viruses. Composite interval mapping in the RIL population identified 17 resistance QTLs associated with the six viruses. Of these, 15 were clustered in specific regions of chr. 2, 3, 6, and 10. It is unknown whether these QTL clusters contain single or multiple virus resistance genes, but the coupling phase linkage of genes conferring resistance to multiple virus diseases in this population could facilitate breeding efforts to develop multi-virus resistant crops.     

Nature of Ribonuclease-resistant Nucleic Acid of Chick Embryo Cells transformed by Schmidt–Ruppin Strain of Rous Sarcoma Virus

THE mode of replication of RNA or RNA-containing tumour viruses is not understood. The recent studies on Rous sarcoma and other RNA-containing oncogenic viruses suggest that the replicative cycle of the RNA of these viruses might not be associated with ribonuclease-resistant structures (double stranded RNAs), but might involve the synthesis of a DNA intermediate specific to viral RNA^1–3. Two groups of workers, however, presented evidence for the presence of a double stranded RNA in 78 Al cell line of rat embryo fibroblasts which had been transformed and chronically infected with the murine sarcoma-leukaemia virus complex (MSV-MLV)^4,5 and it was suggested that the mode of replication of oncogenic viral RNAs was the same as that of non-oncogenic viral RNAs⁴. This apparent discrepancy prompted me to look for ribonuclease-resistant RNA structures in the chick embryo cells transformed by Schmidt-Ruppin Rous sarcoma virus (SR-RSV).