| nucleotide sequence of carnation ringspot dianthovirus rna-2. | rna-2 of carnation ringspot virus (crsv), the type member of the dianthovirus group, has been cdna cloned and sequenced. crsv rna-2 is 1394 nucleotides in length and contains a single open reading frame encoding a 304 amino acid polypeptide of 33.8k. amino acid sequence alignment of this polypeptide with the cell-to-cell movement proteins encoded by rna-2 of red clover necrotic mosaic virus (rcnmv) australian (aus) and czechoslovakian (tpm-34) isolates indicates 59.6% and 55.7% sequence identity ... | 1992 | 1339467 |
| cooperative binding of the red clover necrotic mosaic virus movement protein to single-stranded nucleic acids. | the movement protein of red clover necrotic mosaic dianthovirus was produced in escherichia coli using an expression vector. gel retardation analysis and u.v. cross-linking studies showed that the movement protein bound cooperatively to ssrna and ssdna, but not to dsdna. binding competition experiments established that the movement protein bound to ssrna and ssdna with similar affinities and that the binding was not sequence-specific in the experimental conditions employed. a truncated movement ... | 1992 | 1538188 |
| enzyme-linked immunosorbent assay for detection of red clover necrotic mosaic virus in the host plants. | detection of three isolates of red clover necrotic mosaic virus (rcnmv) representing a, b, and c serotypes was experimentally proved in 18 host plant species by enzyme-linked immunosorbent assay (elisa). in all plant species tested, the homologous serotype reactions showed high selectivity. individual virus serotypes could be reliably detected in the extracts of infected plants only with the homologous igg fraction. group specific detection of rcnmv without serotype determination was possible us ... | 1991 | 1683114 |
| detection of the movement protein of red clover necrotic mosaic virus in a cell wall fraction from infected nicotiana clevelandii plants. | the movement protein of red clover necrotic mosaic virus (rcnmv) was expressed in escherichia coli as a fusion with a maltose-binding protein using the vector pmal-cri and used to produce an antiserum. the rcnmv movement protein was detected in a cell wall fraction obtained from infected nicotiana clevelandii leaf tissue by immunoblotting using the movement protein antiserum. the movement protein could be detected 6 h after inoculation and reached a maximum after 24 h. in contrast, the virus cap ... | 1991 | 1940875 |
| genome structure of tobacco necrosis virus strain a. | an almost complete sequence of the rna genome of tobacco necrosis virus (tnv) strain a has been determined. the genome organization is very similar to that of carnation mottle virus (carmv) and turnip crinkle virus (tcv). the 5'-proximal open reading frame (orf) encodes a 23-kda protein and read-through of its amber codon into the second orf is presumably used for the translation of a 82-kda protein. the third large orf encodes the 30-kda coat protein. two small orfs are located upstream and one ... | 1990 | 2371773 |
| the complete nucleotide sequence and genome organization of red clover necrotic mosaic virus rna-1. | the complete nucleotide sequence of red clover necrotic mosaic virus (rcnmv) rna-1 has been determined. rna-1 is 3889 nucleotides in length with a 5' terminal m7gpppa cap. the rna contains three large open reading frames (orfs): the 5' proximal orf, encoding a 27-kda polypeptide; the internal orf, coding for a 57-kda polypeptide; and the 3' terminal orf, encoding the 37-kda capsid protein. the sequence results confirm in vitro translation of 27-, 50-, and 37-kda products but do not account for t ... | 1989 | 2763465 |
| the nucleotide sequence and gene organization of red clover necrotic mosaic virus rna-2. | red clover necrotic mosaic virus, a member of the dianthovirus group, is characterized by a genome composed of two nonhomologous single-stranded rnas of approximately 4.0 (rna-1) and 1.4 kb (rna-2). the complete nucleotide sequence of the rna-2 has been determined. rna-2 is 1448 nucleotides in length with a 5' terminal m7g cap and no 3' terminal poly-a tail or 5' terminal vpg. an open reading frame beginning at the first initiation codon at nucleotide 80 and ending at nucleotide 1030 has been id ... | 1988 | 3047682 |
| serological properties of certain isolates of red clover necrotic mosaic virus. | | 1969 | 4389112 |
| production and characterization of monoclonal antibodies specific to sweet clover necrotic mosaic virus. | monoclonal antibodies were produced in vitro by fusing mouse myeloma cells p3-x63-agu1 with spleen cells derived from balb/c mice immunized with purified sweet clover necrotic mosaic virus (scnmv). twenty-one out of 47 clones which secreted monoclonal antibodies of high titres against scnmv were injected intraperitoneally into mice previously primed with pristane. the ascites fluid harvested 10 to 14 days later showed a strong and specific anti-scnmv activity in reverse passive haemagglutination ... | 1984 | 6470091 |
| mapping of the red clover necrotic mosaic virus subgenomic rna. | the red clover necrotic mosaic dianthovirus capsid protein is expressed in vivo from a subgenomic rna (sgrna) identical to the 3'-terminal 1.5 kb of rna-1. the 5' leader sequence of the capsid protein sgrna is 62 nucleotides, contains a 14-nucleotide putative promoter sequence homologous to the rna-1 5' terminus, and exhibits a high level of similarity with the tobacco mosaic virus 5' leader translational enhancer element omega. analysis of the rna-1 secondary structure, in the region adjacent t ... | 1996 | 8607270 |
| host-controlled cell-to-cell movement of a hybrid barley stripe mosaic virus expressing a dianthovirus movement protein. | the triple gene block (tgb) of barley stripe mosaic virus (bsmv), coding for viral movement proteins (mps), was replaced by the single mp gene of red clover necrotic mosaic virus (rcnmv). accumulation of the hybrid virus in barley plants (the selective host for bsmv) was reduced compared to bsmv. the hybrid virus induced small necrotic local lesions on chenopodium amaranticolor leaves and did not infect nicotiana clevelandii (the selective host for rcnmv). the hybrid virus accumulated in the ino ... | 1997 | 9268764 |
| mutations in viral movement protein alter systemic infection and identify an intercellular barrier to entry into the phloem long-distance transport system. | viral systemic infection of a plant host involves two processes, cell-to-cell movement and long-distance transport. molecular determinants associated with these two processes were probed by investigating the effects that alanine scanning mutations in the movement protein (mp) of red clover necrotic mosaic virus (rcnmv) had on viral infection in the plant hosts nicotiana edwardsonii, vigna unguiculata (cowpea), and the experimental plant nicotiana benthamiana. plants were inoculated with rcnmv ex ... | 1998 | 9614869 |
| systemic spread of an rna insect virus in plants expressing plant viral movement protein genes. | flock house virus (fhv), a single-stranded rna insect virus, has previously been reported to cross the kingdom barrier and replicate in barley protoplasts and in inoculated leaves of several plant species [selling, b. h., allison, r. f. & kaesberg, p. (1990) proc. natl. acad. sci. usa 87, 434-438]. there was no systemic movement of fhv in plants. we tested the ability of movement proteins (mps) of plant viruses to provide movement functions and cause systemic spread of fhv in plants. we compared ... | 2001 | 11296259 |
| efficient infection of nicotiana benthamiana by tomato bushy stunt virus is facilitated by the coat protein and maintained by p19 through suppression of gene silencing. | tomato bushy stunt virus (tbsv) is one of few rna plant viruses capable of moving systemically in some hosts in the absence of coat protein (cp). tbsv also encodes another protein (p19) that is not required for systemic movement but functions as a symptom determinant in nicotiana benthamiana. here, the role of both cp and p19 in the systemic spread has been reevaluated by utilizing transgenic n. benthamiana plants expressing the movement protein (mp) of red clover necrotic mosaic virus and chime ... | 2002 | 11952121 |
| nucleotide sequence shows that bean leafroll virus has a luteovirus-like genome organization. | the complete nucleotide sequence of the bean leafroll virus (blrv) genomic rna and the termini of its smallest subgenomic rnas were determined to better understand its mechanisms of gene expression and replication and its phylogenetic position within the luteoviridae: the number and placement of open reading frames (orfs) within the blrv genome was luteovirus-like. the nucleotide and predicted amino acid sequences of blrv were most similar to those of soybean dwarf virus (sbdv). phylogenetic ana ... | 2002 | 12075101 |
| molecular analysis of the cucumber leaf spot virus genome. | full-length clones of the genome of the aureusvirus, cucumber leaf spot virus (clsv), have been constructed and infectious t7 polymerase derived synthetic transcripts have been produced. mutational analysis of the genome indicates a role for p84 in viral rna replication, the cp in systemic movement, p27 in viral cell-to-cell movement and p17 in symptom induction. a clsv mutant lacking orfs for the cp, p27 and p17 (clsv yx) was capable of replication and systemic movement in transgenic nicotiana ... | 2003 | 12573495 |
| red clover necrotic mosaic virus replication proteins accumulate at the endoplasmic reticulum. | red clover necrotic mosaic virus (rcnmv) encodes n-terminally overlapping proteins of 27 and 88 kda (p27 and p88) known to be required for replication. green fluorescent protein (gfp) fusions were used to visualize the location of p27 and p88 within nicotiana benthamiana cells. gfp:p27 fusions localized to the endoplasmic reticulum (er), co-localized with er-targeted yellow fluorescent protein and caused membrane restructuring and proliferation. cellular fractionation of virus-inoculated n. bent ... | 2004 | 15016550 |
| structural characterization of an intermolecular rna-rna interaction involved in the transcription regulation element of a bipartite plant virus. | the 34-nucleotide trans-activator (ta) located within the rna-2 of red clover necrotic mosaic virus folds into a simple hairpin. the eight-nucleotide ta loop base pairs with eight complementary nucleotides in the ta binding sequence (tabs) of the capsid protein subgenomic promoter on rna-1 and trans-activates subgenomic rna synthesis. short synthetic oligoribonucleotide mimics of the rna-1 tabs and the rna-2 ta form a weak 1:1 bimolecular complex in vitro with a k(a) of 5.3 x 10(4) m(-1). k(a) d ... | 2004 | 15155850 |
| cap-independent translation mechanism of red clover necrotic mosaic virus rna2 differs from that of rna1 and is linked to rna replication. | the genome of red clover necrotic mosaic virus (rcnmv) in the genus dianthovirus is divided into two rna molecules of rna1 and rna2, which have no cap structure at the 5' end and no poly(a) tail at the 3' end. the 3' untranslated region (3' utr) of rcnmv rna1 contains an essential rna element (3'te-dr1), which is required for cap-independent translation. in this study, we investigated a cap-independent translational mechanism of rna2 using a firefly luciferase (luc) gene expression assay system ... | 2006 | 16571795 |
| in vitro synthesis of minus-strand rna by an isolated cereal yellow dwarf virus rna-dependent rna polymerase requires vpg and a stem-loop structure at the 3' end of the virus rna. | cereal yellow dwarf virus (cydv) rna has a 5'-terminal genome-linked protein (vpg). we have expressed the vpg region of the cydv genome in bacteria and used the purified protein (bvpg) to raise an antiserum which was able to detect free vpg in extracts of cydv-infected oat plants. a template-dependent rna-dependent rna polymerase (rdrp) has been produced from a cydv membrane-bound rna polymerase by treatment with bal 31 nuclease. the rdrp was template specific, being able to utilize templates fr ... | 2006 | 16928757 |
| enhanced resistance and neutralization of defense responses by suppressors of rna silencing. | the effects of transgenic expression of the potato virus y (pvy) hcpro silencing suppressor in tobacco were examined on infection by several viruses. infection by tobacco mosaic virus (tmv) was reduced at 25 degrees c, but not at 33 degrees c. by contrast, systemic infection at 33 degrees c by the tmv expressing green fluorescent protein was promoted by the hcpro. infection by tobacco rattle virus (trv) was restricted to local necrotic lesions by the pvy hcpro. however, this resistance was neutr ... | 2007 | 17617488 |
| use of surface plasmon resonance imaging to study viral rna: protein interactions. | surface plasmon resonance imaging (spri) is an emerging microarray technology that is label-free, rapid and extremely flexible. here the capabilities of spri are demonstrated in results of proof-of-concept experiments detailing a method for studying viral genomic rna:protein interactions in array format. the principal rna is the well-characterized origin of assembly (oas) containing region of tobacco mosaic virus (tmv) rna, whereas the principal protein is the primary subunit for tmv virion asse ... | 2008 | 17875327 |
| translation of red clover necrotic mosaic virus rna in rabbit reticulocyte lysate: identification of the virus coat protein cistron on the larger rna strand of the bipartite genome. | the bipartite genome of red clover necrotic mosaic virus (rcnmv) was translated in nuclease-treated rabbit reticulocyte lysate with the synthesis of three principal translation products of molecular weights 39,000, 36,000, and 34,000. the 39,000 translation product was shown to be rcnmv coat protein by peptide mapping after partial proteolysis and by specific immunoprecipitation. rcnmv coat protein was shown to be coded for on the larger rna (rna-1) of the bipartite genome by hybrid-arrested tra ... | 1983 | 18638867 |
| a viral noncoding rna generated by cis-element-mediated protection against 5'->3' rna decay represses both cap-independent and cap-dependent translation. | positive-strand rna viruses use diverse mechanisms to regulate viral and host gene expression for ensuring their efficient proliferation or persistence in the host. we found that a small viral noncoding rna (0.4 kb), named sr1f, accumulated in red clover necrotic mosaic virus (rcnmv)-infected plants and protoplasts and was packaged into virions. the genome of rcnmv consists of two positive-strand rnas, rna1 and rna2. sr1f was generated from the 3' untranslated region (utr) of rna1, which contain ... | 2008 | 18701589 |
| the red clover necrotic mosaic virus rna-2 encoded movement protein is a second suppressor of rna silencing. | the replication complex of red clover necrotic mosaic virus (rcnmv) has been shown to possess silencing suppression activity. here a newly developed viral-based assay for the identification of silencing suppression activity was used to provide evidence for a second, mechanistically distinct method of silencing suppression provided for by the rcnmv movement protein (mp). this new assay relies on turnip crinkle virus with its capsid protein replaced with green fluorescent protein to act as a repor ... | 2008 | 18838152 |
| the red clover necrotic mosaic virus origin of assembly is delimited to the rna-2 trans-activator. | the bipartite rna genome of red clover necrotic mosaic virus (rcnmv) is encapsidated into icosahedral virions that exist as two populations: i) virions that co-package both genomic rnas and ii) virions packaging multiple copies of rna-2. to elucidate the packaging mechanism, we sought to identify the rcnmv origin of assembly sequence (oas). rcnmv rna-1 cannot package in the absence of rna-2 suggesting that it does not contain an independent packaging signal. a 209 nt rna-2 element expressed from ... | 2009 | 19062064 |
| host-dependent roles of the viral 5' untranslated region (utr) in rna stabilization and cap-independent translational enhancement mediated by the 3' utr of red clover necrotic mosaic virus rna1. | the genome of red clover necrotic mosaic virus (rcnmv) consists of rna1 and rna2, both lacking a cap structure and a poly(a)tail. rna1 has a translational enhancer element (3'te-dr1) in the 3' untranslated region (utr). in this study, we analyzed the roles of 5' and 3' utrs of rna1 in 3'te-dr1-mediated cap-independent translation in cowpea and tobacco by-2 protoplasts using a dual-luciferase (luc) reporter assay system. most mutations introduced into rna1 5' utr in reporter luc mrna abolished or ... | 2009 | 19577782 |
| spontaneous deletion enhances movement of a cucumber necrosis virus based chimera expressing the red clover necrotic mosaic virus movement protein genedagger. | summary the 35-kda movement protein (mp) gene of red clover necrotic mosaic virus (rcnmv) and 3' flanking sequence were inserted in a cucumber necrosis virus (cnv) deletion mutant lacking a large portion of the coding region for the mp. nicotiana benthamiana plants inoculated with chimeric synthetic transcripts of the resulting hybrid cdna clone (m5/rm2) developed both local and systemic symptoms and accumulated high levels of chimeric viral rna. reverse transcriptase polymerase chain reaction ( ... | 2001 | 20572988 |
| traffic jam on the cellular secretory pathway generated by a replication protein from a plant rna virus. | although positive-strand rna [(+)rna] viruses have a limited coding capacity, they can replicate efficiently in host cells because of their ability to use host-derived proteins, membranes, lipids, and metabolites, and to rewire cellular trafficking pathways. previously, we showed that a plant rna virus, the red clover necrotic mosaic virus (rcnmv), hijacked arf1 and sar1, which are small gtpases that regulate the biogenesis of copi and copii vesicles, respectively, for viral rna replication. the ... | 2014 | 24714629 |
| phosphatidic acid produced by phospholipase d promotes rna replication of a plant rna virus. | eukaryotic positive-strand rna [(+)rna] viruses are intracellular obligate parasites replicate using the membrane-bound replicase complexes that contain multiple viral and host components. to replicate, (+)rna viruses exploit host resources and modify host metabolism and membrane organization. phospholipase d (pld) is a phosphatidylcholine- and phosphatidylethanolamine-hydrolyzing enzyme that catalyzes the production of phosphatidic acid (pa), a lipid second messenger that modulates diverse intr ... | 2015 | 26020241 |
| harnessing host ros-generating machinery for the robust genome replication of a plant rna virus. | as sessile organisms, plants have to accommodate to rapid changes in their surrounding environment. reactive oxygen species (ros) act as signaling molecules to transduce biotic and abiotic stimuli into plant stress adaptations. it is established that a respiratory burst oxidase homolog b of nicotiana benthamiana (nbrbohb) produces ros in response to microbe-associated molecular patterns to inhibit pathogen infection. plant viruses are also known as causative agents of ros induction in infected p ... | 2017 | 28154139 |
| noncoding rnas of plant viruses and viroids: sponges of host translation and rna interference machinery. | noncoding sequences in plant viral genomes are well-known to control viral replication and gene expression in cis. however, plant viral and viroid noncoding (nc)rna sequences can also regulate gene expression acting in trans, often acting like 'sponges' that bind and sequester host cellular machinery to favor viral infection. noncoding sequences of small subgenomic (sg)rnas of barley yellow dwarf virus (bydv) and red clover necrotic mosaic virus (rcnmv) contain a cap-independent translation elem ... | 2016 | 26900786 |
| red clover necrotic mosaic virus infectious transcripts synthesized in vitro. | the red clover necrotic mosaic dianthovirus (rcnmv) genome is split between two essentially nonhomologous ssrnas of 3.9 kb (rna-1) and 1.45 kb (rna-2) which are each capped at the 5' terminus with m7gpppa. cdna clones short of full length by several nucleotides at both termini have been generated to both rnas. oligonucleotide-directed mutagenesis was employed to generate a series of rna-1 and -2 transcription vectors in which the bacteriophage t7 rna polymerase promoter was fused to full-length ... | 1991 | 2024474 |
| development of abamectin loaded plant virus nanoparticles for efficacious plant parasitic nematode control. | plant parasitic nematodes are one of the world's major agricultural pests, causing in excess of $157 billion in worldwide crop damage annually. abamectin (abm) is a biological pesticide with a strong activity against a wide variety of plant parasitic nematodes. however, abm's poor mobility in the soil compromises its nematicide performance because of the limited zone of protection surrounding the growing root system of the plant. in this study, we manipulated abm's soil physical chemistry by enc ... | 2015 | 25906360 |
| a spontaneous red clover necrotic mosaic virus mutant with a truncated movement protein. | a spontaneous red clover necrotic mosaic virus mutant, tpm-341, was isolated by multiple passage of czechoslovakian isolate tpm-34 in beans, followed by three cycles of single lesion isolation in cowpea and in chenopodium quinoa. the symptoms induced in cowpea by tpm-34 and tpm-341 differed. tpm-34 gave rise to chlorotic lesions which expanded with time, often becoming confluent with adjacent lesions, and developed necrotic margins; the plants became systemically infected. tpm-341 induced necrot ... | 1991 | 1875192 |
| proteolytic processing of cmpp36, a protein from the cytochrome b(5) reductase family, is required for entry into the phloem translocation pathway. | cucurbita maxima (pumpkin) phloem sap contains a 31 kda protein that cross-reacts with antibodies directed against the red clover necrotic mosaic virus movement protein (rcnmv mp). microsequence data from phloem-purified 31 kda protein were used to isolate a complementary dna: the open reading frame encodes a 36 kda protein belonging to the cytochrome b(5) reductase (cb5r) family; the gene was termed cmpp36. western analyses established that cmpp36, rcnmv mp and cmpp16 (xoconostle-cázares et al. ... | 2000 | 11135108 |
| a single amino acid mutation in the carnation ringspot virus capsid protein allows virion formation but prevents systemic infection. | a carnation ringspot virus (crsv) variant (1.26) was identified that accumulates virions but is incapable of forming a systemic infection. the 1.26 capsid protein gene possesses a ser-->pro mutation at amino acid 282. conversion of 1.26 amino acid 282 to ser restored systemic infection, while the reciprocal mutation in wild-type crsv abolished systemic infection. similar mutations introduced into the related red clover necrotic mosaic virus capsid protein gene failed to induce the packaging but ... | 2001 | 11533217 |
| sv40 vp1 major capsid protein in its self-assembled form allows vp1 pentamers to coat various types of artificial beads in vitro regardless of their sizes and shapes. | the icosahedral capsid structure of simian virus 40 (diameter, 45 nm) consists of 72 pentameric subunits, with each subunit formed by five vp1 molecules. electron microscopy, immuno-gold labeling, and ζ-potential analysis showed that purified recombinant vp1 pentamers covered polystyrene beads measuring 100, 200, and 500 nm in diameter, as well as silica beads. in addition to covering spherical beads, vp1 pentamers covered cubic magnetite beads, as well as the distorted surface structures of lip ... | 2015 | 28435806 |
| roles of superoxide anion and hydrogen peroxide during replication of two unrelated plant rna viruses in nicotiana benthamiana. | reactive oxygen species (ros), including superoxide anion (o2(-)), hydrogen peroxide (h2o2), and hydroxyl radical, act as signaling molecules to transduce biotic and abiotic stimuli into stress adaptations in plants. a respiratory burst oxidase homolog b of nicotiana benthamiana (nbrbohb) is responsible for o2(-) production to inhibit pathogen infection during plant innate immunity. rboh-derived o2(-) can be immediately converted into h2o2 by the action of superoxide dismutase. interestingly, we ... | 2017 | 28594275 |
| requirement for eukaryotic translation initiation factors in cap-independent translation differs between bipartite genomic rnas of red clover necrotic mosaic virus. | the bipartite genomic rnas of red clover necrotic mosaic virus (rcnmv) lack a 5' cap and a 3' poly(a) tail. rna1 encodes viral replication proteins, and rna2 encodes a movement protein (mp). these proteins are translated in a cap-independent manner. we previously identified two cis-acting rna elements that cooperatively recruit eukaryotic translation initiation factor (eif) complex eif4f or eifiso4f to rna1. such cis-acting rna elements and host factors have not been identified in rna2. here we ... | 2017 | 28646650 |
| gapdh--a recruits a plant virus movement protein to cortical virus replication complexes to facilitate viral cell-to-cell movement. | the formation of virus movement protein (mp)-containing punctate structures on the cortical endoplasmic reticulum is required for efficient intercellular movement of red clover necrotic mosaic virus (rcnmv), a bipartite positive-strand rna plant virus. we found that these cortical punctate structures constitute a viral replication complex (vrc) in addition to the previously reported aggregate structures that formed adjacent to the nucleus. we identified host proteins that interacted with rcnmv m ... | 2014 | 25411849 |
| loading and release mechanism of red clover necrotic mosaic virus derived plant viral nanoparticles for drug delivery of doxorubicin. | loading and release mechanisms of red clover necrotic mosaicvirus (rcnmv) derived plant viral nanoparticle (pvn) are shown for controlled delivery of the anticancer drug, doxorubicin (dox). previous studies demonstrate that rcnmv's structure and unique response to divalent cation depletion and re-addition enables dox infusion to the viral capsid through a pore formation mechanism. however, by controlling the net charge of rcnmv outer surface and accessibility of rcnmv interior cavity, tunable re ... | 2014 | 25098668 |
| the red clover necrotic mosaic virus capsid protein n-terminal amino acids possess specific rna binding activity and are required for stable virion assembly. | the red clover necrotic mosaic virus (rcnmv) bipartite rna genome is packaged into two virion populations containing either rna-1 and rna-2 or multiple copies of rna-2 only. to understand this distinctive packaging scheme, we investigated the rna-binding properties of the rcnmv capsid protein (cp). maltose binding protein-cp fusions exhibited the highest binding affinities for rna probes containing the rna-2 trans-activator or the 3' non-coding region from rna-1. other viral and non-viral rna pr ... | 2013 | 23747688 |
| new insight into the structure of rna in red clover necrotic mosaic virus and the role of divalent cations revealed by small-angle neutron scattering. | red clover necrotic mosaic virus (rcnmv) is a 36-nm-diameter, t = 3 icosahedral plant virus with a genome that is split between two single-stranded rna molecules of approximately 3.9 kb and 1.5 kb, as well as a 400-nucleotide degradation product. the structure of the virus capsid and its response to removing ca(2+) and mg(2+) was previously studied by cryo-electron microscopy (cryo-em) (sherman et al. j virol 80:10395-10406, 2006) but the structure of the rna was only partially resolved in that ... | 2013 | 23483344 |
| polymeric systems incorporating plant viral nanoparticles for tailored release of therapeutics. | therapeutic polylactide (pla) nanofibrous matrices are fabricated by incorporating plant viral nanoparticles (pvns) infused with fluorescent agents ethidium bromide (etbr) and rhodamine (rho), and cancer therapeutic doxorubicin (dox). the native virus, red clover necrotic mosaic virus (rcnmv), reversibly opens and closes upon exposure to the appropriate environmental stimuli. infusing rcnmv with small molecules allows the incorporation of pvn(active) into fibrous matrices via two methods: direct ... | 2013 | 23335438 |
| avidity-mediated virus separation using a hyperthermophilic affinity ligand. | immunoaffinity separation of large multivalent species such as viruses is limited by the stringent elution conditions necessary to overcome their strong and highly avid interaction with immobilized affinity ligands on the capture surface. here we present an alternate strategy that harnesses the avidity effect to overcome this limitation. red clover necrotic mosaic virus (rcnmv), a plant virus relevant to drug delivery applications, was chosen as a model target for this study. an rcnmv binding pr ... | 2013 | 23125177 |
| adp ribosylation factor 1 plays an essential role in the replication of a plant rna virus. | eukaryotic positive-strand rna viruses replicate using the membrane-bound replicase complexes, which contain multiple viral and host components. virus infection induces the remodeling of intracellular membranes. virus-induced membrane structures are thought to increase the local concentration of the components that are required for replication and provide a scaffold for tethering the replicase complexes. however, the mechanisms underlying virus-induced membrane remodeling are poorly understood. ... | 2013 | 23097452 |
| differential roles of hsp70 and hsp90 in the assembly of the replicase complex of a positive-strand rna plant virus. | assembly of viral replicase complexes of eukaryotic positive-strand rna viruses is a regulated process: multiple viral and host components must be assembled on intracellular membranes and ordered into quaternary complexes capable of synthesizing viral rnas. however, the molecular mechanisms underlying this process are poorly understood. in this study, we used a model virus, red clover necrotic mosaic virus (rcnmv), whose replicase complex can be detected readily as the 480-kda functional protein ... | 2012 | 22933272 |
| identification of domains in p27 auxiliary replicase protein essential for its association with the endoplasmic reticulum membranes in red clover necrotic mosaic virus. | positive-strand rna viruses require host intracellular membranes for replicating their genomic rnas. in this study, we determined the domains and critical amino acids in p27 of red clover necrotic mosaic virus (rcnmv) required for its association with and targeting of er membranes in nicotiana benthamiana plants using a c-terminally gfp-fused and biologically functional p27. confocal microscopy and membrane-flotation assays using an agrobacterium-mediated expression system showed that a stretch ... | 2012 | 22898643 |
| poly(a)-binding protein facilitates translation of an uncapped/nonpolyadenylated viral rna by binding to the 3' untranslated region. | viruses employ an alternative translation mechanism to exploit cellular resources at the expense of host mrnas and to allow preferential translation. plant rna viruses often lack both a 5' cap and a 3' poly(a) tail in their genomic rnas. instead, cap-independent translation enhancer elements (cites) located in the 3' untranslated region (utr) mediate their translation. although eukaryotic translation initiation factors (eifs) or ribosomes have been shown to bind to the 3'cites, our knowledge is ... | 2012 | 22593149 |
| the red clover necrotic mosaic virus capsid protein n-terminal lysine-rich motif is a determinant of symptomatology and virion accumulation. | the interaction between viral capsid protein (cp) and its cognate viral rna modulates many steps in the virus infection cycle, such as replication, translation and assembly. the n-terminal 50 amino acids of the red clover necrotic mosaic virus (rcnmv) cp are rich in basic residues (especially lysine) and are essential for the core functions of the cp, namely rna binding and virion assembly. to further elucidate additional biological roles for these basic residues, a series of alanine substitutio ... | 2012 | 22292426 |
| mass spectrometric detection of targeting peptide bioconjugation to red clover necrotic mosaic virus. | plant virus nanoparticle (pvn) formulations constructed from red clover necrotic mosaic virus by drug infusion and targeting peptide conjugation can be employed as drug delivery tools. in this investigation, we studied the cross-linked structures formed by application of sulfosuccinimidyl-4-(n-maleimidomethyl) cyclohexane-1-carboxylate (ssmcc) and succinimidyl-[(n-maleimidopropionamido)-hexaethylene glycol] ester (smpeg) as heterobifunctional linkers in the bioconjugation process. the plant viru ... | 2011 | 21854060 |
| a long-distance rna-rna interaction plays an important role in programmed -1 ribosomal frameshifting in the translation of p88 replicase protein of red clover necrotic mosaic virus. | programmed -1 ribosomal frameshifting (-1 prf) is one viral translation strategy to express overlapping genes in positive-strand rna viruses. red clover necrotic mosaic virus (rcnmv) uses this strategy to express its replicase component protein p88. in this study, we used a cell-free translation system to map cis-acting rna elements required for -1 prf. our results show that a small stem-loop structure adjacent to the cap-independent translation element in the 3' untranslated region (utr) of rcn ... | 2011 | 21703656 |
| identification of amino acids in auxiliary replicase protein p27 critical for its rna-binding activity and the assembly of the replicase complex in red clover necrotic mosaic virus. | the specific recognition of genomic rnas by viral replicase proteins is a key regulatory step during the early replication process in positive-strand rna viruses. in this study, we characterized the rna-binding activity of the auxiliary replicase protein p27 of red clover necrotic mosaic virus (rcnmv), which has a bipartite genome consisting of rna1 and rna2. aptamer pull-down assays identified the amino acid residues of p27 involved in its specific interaction with rna2. the rna-binding activit ... | 2011 | 21440279 |
| viruses as nanomaterials for drug delivery. | virus delivery vectors are one among the many nanomaterials that are being developed as drug delivery materials. this chapter focuses on methods utilizing plant virus nanoparticles (pvns) synthesized from the red clover necrotic mosaic virus (rcnmv). a successful vector must be able to effectively carry and subsequently deliver a drug cargo to a specific target. in the case of the pvns, we describe two types of ways cargo can be loaded within these structures: encapsidation and infusion. several ... | 2011 | 21424452 |
| viral cell-to-cell movement requires formation of cortical punctate structures containing red clover necrotic mosaic virus movement protein. | movement protein (mp) of red clover necrotic mosaic virus (rcnmv) forms punctate structures on the cortical endoplasmic reticulum (er) of nicotiana benthamiana cells, which are associated with viral rna1 replication (kaido et al., virology 395, 232-242. 2009). we investigated the significance of er-targeting by mp during virus movement from cell to cell, by analyzing the function of a series of mps with varying length deletions at their c-terminus, either fused or not fused with green fluorescen ... | 2011 | 21377183 |
| the red clover necrotic mosaic virus capsid as a multifunctional cell targeting plant viral nanoparticle. | multifunctional nanoparticles hold promise as the next generation of therapeutic delivery and imaging agents. nanoparticles comprising many types of materials are being tested for this purpose, including plant viral capsids. it has been found that red clover necrotic mosaic virus (rcnmv) can be loaded with significant amounts of therapeutic molecules with molecular weights of 600 or even greater. formulation of rcnmv into a plant viral nanoparticle (pvn) involves the loading of cargo and attachm ... | 2011 | 21126069 |
| crystallization and preliminary x-ray diffraction analysis of red clover necrotic mosaic virus. | red clover necrotic mosaic virus (rcnmv) is a species that belongs to the tombusviridae family of plant viruses with a t = 3 icosahedral capsid. rcnmv virions were purified and were crystallized for x-ray analysis using the hanging-drop vapor-diffusion method. self-rotation functions and systematic absences identified the space group as i23, with two virions in the unit cell. the crystals diffracted to better than 4 å resolution but were very radiation-sensitive, causing rapid decay of the high- ... | 2010 | 21045294 |
| template recognition mechanisms by replicase proteins differ between bipartite positive-strand genomic rnas of a plant virus. | recognition of rna templates by viral replicase proteins is one of the key steps in the replication process of all rna viruses. however, the mechanisms underlying this phenomenon, including primary rna elements that are recognized by the viral replicase proteins, are not well understood. here, we used aptamer pulldown assays with membrane fractionation and protein-rna coimmunoprecipitation in a cell-free viral translation/replication system to investigate how viral replicase proteins recognize t ... | 2011 | 20980498 |
| interactions between p27 and p88 replicase proteins of red clover necrotic mosaic virus play an essential role in viral rna replication and suppression of rna silencing via the 480-kda viral replicase complex assembly. | red clover necrotic mosaic virus (rcnmv), a positive-sense rna virus with a bipartite genome, encodes p27 and p88 replicase proteins that are required for viral rna replication and suppression of rna silencing. in this study, we identified domains in p27 and p88 responsible for their protein-protein interactions using in vitro pull-down assays with the purified recombinant proteins. coimmunoprecipitation analysis in combination with blue-native polyacrylamide gel electrophoresis using mutated p2 ... | 2010 | 20828775 |
| a y-shaped rna structure in the 3' untranslated region together with the trans-activator and core promoter of red clover necrotic mosaic virus rna2 is required for its negative-strand rna synthesis. | red clover necrotic mosaic virus (rcnmv) is a positive-strand rna virus with a bipartite genome. rna1 encodes n-terminally overlapping replication proteins, p27 and p88. rna2 is replicated efficiently by the replication proteins supplied in trans, whereas rna1 needs p88 preferentially in cis for its replication. cis-acting elements required for rna2 replication have been mapped to the 3' terminal stem-loop structure conserved between rna1 and rna2, and to the protein-coding region including the ... | 2010 | 20561661 |
| identification and characterization of the 480-kilodalton template-specific rna-dependent rna polymerase complex of red clover necrotic mosaic virus. | replication of positive-strand rna viruses occurs through the assembly of membrane-associated viral rna replication complexes that include viral replicase proteins, viral rna templates, and host proteins. red clover necrotic mosaic virus (rcnmv) is a positive-strand rna plant virus with a genome consisting of rna1 and rna2. the two proteins encoded by rna1, a 27-kda protein (p27) and an 88-kda protein containing an rna-dependent rna polymerase (rdrp) motif (p88), are essential for rcnmv rna repl ... | 2010 | 20375154 |
| endoplasmic reticulum targeting of the red clover necrotic mosaic virus movement protein is associated with the replication of viral rna1 but not that of rna2. | red clover necrotic mosaic virus (rcnmv) is a positive-strand rna virus with a bipartite genome. the movement protein (mp) encoded by rna2 is essential for viral movement. to obtain further insights into the viral movement mechanism, subcellular localizations of rcnmv mp fused with green fluorescent protein (mp:gfp) were examined in nicotiana benthamiana epidermal cells and protoplasts. the mp:gfp expressed from the recombinant virus first appeared in the cell wall and subsequently was observed ... | 2009 | 19819513 |
| three discontinuous loop nucleotides in the 3' terminal stem-loop are required for red clover necrotic mosaic virus rna-2 replication. | the genome of red clover necrotic mosaic virus (rcnmv) consists of positive-sense, single-stranded rna-1 and rna-2. the 29 nucleotides at the 3' termini of both rnas are nearly identical and are predicted to form a stable stem-loop (sl) structure, which is required for rcnmv rna replication. here we performed a systematic mutagenesis of the rna-2 3' sl to identify the nucleotides critical for replication. infectivity and rna replication assays indicated that the secondary structure of the 3' sl ... | 2009 | 19733887 |
| targeting cancer with 'smart bombs': equipping plant virus nanoparticles for a 'seek and destroy' mission. | this article discusses plant virus nanoparticles as a weapon in the war on cancer. the successes and failures of numerous nanoparticle strategies are discussed as a background to consideration of the plant virus nanoparticle approach. to have therapeutic benefit, the advantages of the targeted nanoparticle must outweigh the problems of colloidal stability, uptake by the reticuloendothelial system as well as the requirement for clearance from the body. biodegradable nanoparticles are considered t ... | 2009 | 19572822 |
| infusion of dye molecules into red clover necrotic mosaic virus. | the red clover necrotic mosaic virus capsid is utilized to package and release molecules through reversible depletion and re-addition of divalent cations. | 2008 | 18401897 |
| cis-preferential requirement of a -1 frameshift product p88 for the replication of red clover necrotic mosaic virus rna1. | the genome of red clover necrotic mosaic virus (rcnmv) consists of rna1 and rna2. rna1 encodes n-terminally overlapping replication proteins, p27 and p88, which are translated in a cap-independent manner. the 3' untranslated region of rna1 contains rna elements essential for cap-independent translation and negative-strand rna synthesis. in this study, we investigated how p27 and p88 were engaged in the replication of rcnmv genomic rnas by using dna vectors or in vitro transcribed rnas in protopl ... | 2008 | 18308359 |
| cis-acting core rna elements required for negative-strand rna synthesis and cap-independent translation are separated in the 3'-untranslated region of red clover necrotic mosaic virus rna1. | the genome of red clover necrotic mosaic virus (rcnmv) is positive-sense and divided into rna1 and rna2. rna1 has a translation enhancer element (3' te-dr1) in the 3' untranslated region (utr) that substitutes for a 5' cap. in this study, we determined the regions required for cap-independent translation and rna synthesis in the 3' utr of rna1 using a cell-free extract of evacuolated by-2 protoplasts (byl) and by an assay in by-2 protoplasts. the use of capped viral rna transcripts in the byl sy ... | 2007 | 17727911 |
| encapsidation of nanoparticles by red clover necrotic mosaic virus. | icosahedral virus capsids demonstrate a high degree of selectivity in packaging cognate nucleic acid genome components during virion assembly. the 36 nm icosahedral plant virus red clover necrotic mosaic virus (rcnmv) packages its two genomic ssrnas via a specific capsid protein (cp) genomic rna interaction. a 20-nucleotide hairpin structure within the genomic rna-2 hybridizes with rna-1 to form a bimolecular complex, which is the origin of assembly (oas) in rcnmv that selectively recruits and o ... | 2007 | 17705477 |
| removal of divalent cations induces structural transitions in red clover necrotic mosaic virus, revealing a potential mechanism for rna release. | the structure of red clover necrotic mosaic virus (rcnmv), an icosahedral plant virus, was resolved to 8.5 a by cryoelectron microscopy. the virion capsid has prominent surface protrusions and subunits with a clearly defined shell and protruding domains. the structures of both the individual capsid protein (cp) subunits and the entire virion capsid are consistent with other species in the tombusviridae family. within the rcnmv capsid, there is a clearly defined inner cage formed by complexes of ... | 2006 | 16920821 |
| controlled encapsidation of gold nanoparticles by a viral protein shell. | icosahedral virus capsids demonstrate a high degree of selectivity in packaging cognate nucleic acid components during assembly. this packaging specificity, when integrated as part of a nanotechnological protocol, has the potential to encapsidate a wide array of foreign materials for delivery of therapeutics or biosensors into target cells. red clover necrotic mosaic virus (rcnmv) exclusively packages two genomic ssrnas initiated by a specific protein:rna interaction between the rcnmv coat prote ... | 2006 | 16594649 |
| the genomic rna packaging scheme of red clover necrotic mosaic virus. | red clover necrotic mosaic virus (rcnmv) is a small icosahedral plant virus with a bipartite rna genome. while the rcnmv genome consists of two rnas, it has not been definitively established whether these rnas are co-packaged into a single virion or packaged individually into separate virions. biochemical evidence exists to support both hypotheses. to determine the genomic rna complement within rcnmv, virions were subjected to heat treatments and uv crosslinking. a stable rna-1:rna-2 heterodimer ... | 2006 | 16297955 |
| a plant rna virus suppresses rna silencing through viral rna replication. | rna interference (rnai) is a post-transcriptional gene-regulatory mechanism that operates in many eukaryotes. rnai is induced by double-stranded rna (dsrna) and is mainly involved in defence against transposons and viruses. to counteract rnai, viruses have rnai suppressors. here we show a novel mechanism of rnai suppression by a plant virus red clover necrotic mosaic virus (rcnmv). to suppress rnai, rcnmv needs multiple viral components, which include viral rnas and putative rna replicase protei ... | 2005 | 16096641 |
| cell wall localization of red clover necrotic mosaic virus movement protein is required for cell-to-cell movement. | the red clover necrotic mosaic virus movement protein (mp) is essential for cell-to-cell movement. eight previously characterized alanine-scanning mutants of the mp were fused to the green fluorescent protein (gfp) and expressed from viral infectious transcripts. inoculated plants were assayed for movement and intracellular accumulation of mp by confocal laser-scanning microscopy. a strict correlation was observed between the targeting to the cell wall (presumably the plasmodesmata) and cell-to- ... | 2005 | 15708588 |
| the red clover necrotic mosaic virus rna2 trans-activator is also a cis-acting rna2 replication element. | the expression of the coat protein gene requires rna-mediated trans-activation of subgenomic rna synthesis in red clover necrotic mosaic virus (rcnmv), the genome of which consists of two positive-strand rnas, rna1 and rna2. the trans-acting rna element required for subgenomic rna synthesis from rna1 has been mapped previously to the protein-coding region of rna2, whereas rna2 is not required for the replication of rna1. in this study, we investigated the roles of the protein-coding region in rn ... | 2005 | 15613326 |
| a sobemovirus coat protein gene complements long-distance movement of a coat protein-null dianthovirus. | red clover necrotic mosaic virus (rcnmv; genus dianthovirus) and turnip rosette virus (trov; genus sobemovirus) are taxonomically and ecologically distinct plant viruses. in addition, the two genera differ in the role of coat protein (cp) in cell-to-cell movement. however, both are small icosahedral viruses requiring cp for systemic movement in the host vasculature. here, we show that the trov cp gene is capable of facilitating the vascular movement of a dianthovirus. substitution of the rcnmv c ... | 2004 | 15527845 |
| cap-independent translational enhancement by the 3' untranslated region of red clover necrotic mosaic virus rna1. | red clover necrotic mosaic virus (rcnmv) is a member of the genus dianthovirus and has a bipartite positive-sense genomic rna with 3' ends that are not polyadenylated. in this study, we show that both genomic rna1 and rna2 lack a 5' cap structure and that uncapped in vitro transcripts of rcnmv rna1 replicated to a level comparable to that for capped transcripts in cowpea protoplasts. because the 5' cap and 3' poly(a) tail play important roles in the translation of many eukaryotic mrnas, genomic ... | 2003 | 14581548 |
| the 3'-untranslated region of rna1 as a primary determinant of temperature sensitivity of red clover necrotic mosaic virus canadian strain. | red clover necrotic mosaic virus canadian strain (rcnmv-can) induces symptoms on host plants at 17 degrees c, but not at 25 degrees c. we investigated the temperature sensitivity of rcnmv-can in nicotiana benthamiana plants and protoplasts using infectious transcripts of genomic rnas 1 and 2. viral rnas accumulated in both inoculated and noninoculated leaves at 17 degrees c, whereas no viral rnas were detected at 25 degrees c in either inoculated or noninoculated leaves. similar temperature sens ... | 2002 | 11886252 |
| mutational analysis of cis-acting sequences in the 3'- and 5'-untranslated regions of rna2 of red clover necrotic mosaic virus. | mutational analysis has been used to investigate cis-acting sequences in the 3'- and 5'-untranslated regions of red clover necrotic mosaic virus rna2 required for replication in the presence of wild-type rna1. deletion of a sequence near the 3' end of rna2, which is potentially capable of forming a stable stem-loop structure, abolished the ability of rna2 to replicate in nicotiana clevelandii protoplasts, as judged by the failure to detect production of either the positive or negative rna strand ... | 1999 | 9887324 |
| rna-mediated trans-activation of transcription from a viral rna. | the red clover necrotic mosaic virus genome is composed of two single-stranded rna components, rna-1 and rna-2. the viral capsid protein is translated from a subgenomic rna (sgrna) that is transcribed from genomic rna-1. here, a 34-nucleotide sequence in rna-2 is shown to be required for transcription of sgrna. mutations that prevent base-pairing between the rna-1 subgenomic promoter and the 34-nucleotide trans-activator prevent expression of a reporter gene. a model is proposed in which direct ... | 1998 | 9694655 |
| virion formation is required for the long-distance movement of red clover necrotic mosaic virus in movement protein transgenic plants. | the red clover necrotic mosaic dianthovirus (rcnmv) genome is split between two single-stranded rna species. the polycistronic rna-1 encodes the viral rna polymerase and capsid protein (cp) and the monocistronic rna-2 encodes the 35 kda cell-to-cell movement protein (mp). nicotiana benthamiana plants transformed with the rcnmv mp gene were generated. when inoculated onto the mp transgenic plants, cell-to-cell movement of rna-1 occurred at a rate similar to wild-type virus. however, long-distance ... | 1995 | 7571430 |
| serological typing of red clover necrotic mosaic virus isolates by enzyme-linked immunosorbent assay. | enzyme-linked immunosorbent assay (elisa) enabled a more precise serological typing of isolates of red clover necrotic mosaic virus (rcnmv) originating from slovak and czech republics, poland, sweden and great britain. we found 5 isolates of serotype a, 14 isolates of serotype b, and 6 isolates of serotype c. some isolates represented mixtures of serotypes, namely a+b (1 isolate), a+c (1), b+c (7), and a+b+c (5). elisa was found to be a more suitable method of serotype identification of rcnmv is ... | 1994 | 7879713 |
| identification and analysis of the site of -1 ribosomal frameshifting in red clover necrotic mosaic virus. | the genomic rna-1 of red clover necrotic mosaic dianthovirus (rcnmv) contains the heptanucleotide ggauuuu that precedes the termination codon of the 5' proximal p27 open reading frame (orf). this heptanucleotide is followed by a sequence with the potential to form a stable, complex secondary structure. translation of rna-1 is postulated to utilize a -1 ribosomal frameshifting mechanism to express the 88-kda viral rna polymerase. using site-directed mutagenesis together with cell-free translation ... | 1994 | 8178444 |
| cell-to-cell trafficking of macromolecules through plasmodesmata potentiated by the red clover necrotic mosaic virus movement protein. | direct evidence is presented for cell-to-cell trafficking of macromolecules via plasmodesmata in higher plants. the fluorescently labeled 35-kd movement protein of red clover necrotic mosaic virus (rcnmv) trafficked rapidly from cell to cell when microinjected into cowpea leaf mesophyll cells. furthermore, this protein potentiated rapid cell-to-cell trafficking of rcnmv rna, but not dna. electron microscopic studies demonstrated that the 35-kd movement protein does not unfold the rcnmv rna molec ... | 1993 | 12271056 |
| alanine scanning mutagenesis of a plant virus movement protein identifies three functional domains. | alanine scanning mutagenesis was performed on the red clover necrotic mosaic virus (rcnmv) movement protein (mp), and 12 mutants were assayed in vitro for rna binding characteristics and in vivo for their ability to potentiate rcnmv cell-to-cell movement. the mutant phenotypes that were identified in vitro and in vivo suggest both that cooperative rna binding is not necessary for cell-to-cell movement in vivo and that only a fraction of the wild-type rna binding may be required. the mp mutants d ... | 1993 | 8400873 |
| synthesis of the putative red clover necrotic mosaic virus rna polymerase by ribosomal frameshifting in vitro. | the red clover necrotic mosaic virus (rcnmv) genome is split between two single-stranded rna species termed rna-1 and rna-2. rna-1 directs the synthesis of 88-kda (p88), 57-kda (p57), 37-kda (p37), and 27-kda (p27) polypeptides and rna-2 a 35-kda (p35) polypeptide in vitro. the coding order of the rna-1 products was determined to be 5'-p27-p57-p37-3'. antibodies to synthetic peptides representing the carboxyl terminal portions of p27 and p57 immunoprecipitated their respective polypeptides in ad ... | 1993 | 8438566 |
| the roles of the red clover necrotic mosaic virus capsid and cell-to-cell movement proteins in systemic infection. | the red clover necrotic mosaic dianthovirus (rcnmv) genome is split between two single-stranded rna species termed rna-1 and rna-2. rna-2 is required for infection of whole plants but is dispensable for infection and virion formation in protoplasts. we have used full-length cdna clones of rna-1 and -2 from which infectious in vitro transcripts can be derived to construct a number of mutations in the rna-1 encoded capsid protein and the rna-2 encoded cell-to-cell movement protein genes. the capsi ... | 1993 | 8517020 |
| a comparative study of the rna-2 nucleotide sequences of two sweet clover necrotic mosaic virus strains. | the nucleotide sequences of the rna-2s of two strains of sweet clover necrotic mosaic virus (scnmv-38 and -59) have been determined. the rna-2s of scnmv-38 and -59 consist of 1446 and 1449 nucleotides, respectively, and both contain one major open reading frame (orf) which potentially can encode polypeptides of 326 amino acid residues (about 36.5k), designated sc38p2 and sc59p2, respectively. the nucleotide sequences of scnmv-38 and -59 rna-2s show 93.2% similarity, and the amino acid sequences ... | 1992 | 1402823 |
| nucleotide sequence of rna 2 of a czechoslovakian isolate of red clover necrotic mosaic virus. | the complete nucleotide sequence (1448 nucleotides) of rna 2 of a czechoslovakian isolate tpm-34 of red clover necrotic mosaic virus (rcnmv-tpm-34) has been determined. the sequence contained one major open reading frame (orf) with the potential to encode a protein of 326 amino acids (mr 35755), designated p2. the nucleotide sequence of rna 2 of rcnmv-tpm-34 and the previously published sequence of rna 2 of an australian isolate of the virus (rcnmv-aus) were 83% identical and there was 80% amino ... | 1991 | 1990066 |
| determination of serotypes of red clover necrotic mosaic virus by enzyme-linked immunosorbent assay. | the direct double antibody sandwich (das) type of enzyme-linked immunosorbent assay (elisa) was used to determine the degree of serological and antigenic differences, among the three serotypes (a, b, c) of red clover necrotic mosaic virus (rcnmv). homologous and heterologous antibody titres in the used iggs to isolates tpm34 (serotype a), tpm48 (serotype b) and isolate no. 6 (serotype c) as determined by elisa were 100- to 200-fold higher than by ring precipitation test. intensity of homologous ... | 1990 | 1705747 |
| isoelectric points of red clover necrotic mosaic virus serotypes. | isoelectric focusing in agarose gel was used to determine the isoelectric points of red clover necrotic mosaic virus serotypes. the isoelectric points of serotypes a, b and c were in the ph ranges from 5.03-5.06, from 4.82-4.84 and from 4.60-4.62, respectively. | 1984 | 6143503 |
| serotypes of red clover necrotic mosaic virus. iii. immunoelectrophoresis under different conditions. | the effects of 4 buffers at different ph and of different molarities was tested on the immunoelectrophoresis of representatives of three serotypes and their natural mixtures of red clover necrotic mosaic virus (rcnmv). no substantial differences were observed with buffers at ph 7.2, 8.0 and 8.6. the representatives of the three rcnmv serotypes showed different mobilities in 0.1 and 0.01 mol/l ionic buffers in the agarose gel from cathode to anode. in nonionic 0.01-0.1 mol/l n-2-hydroxyethylpiper ... | 1984 | 6143502 |
| serotypes of red clover necrotic mosaic virus. i. characterization of three serotypes. | three serotypes (a, b and c) were distinguished based on serological differences between isolates of red clover necrotic mosaic virus (rcnmv). isolate tpm34, representative of serotype a, induced the formation of antibody only against homologous antigen. by contrast, isolate tpm48, representative of serotype b, induced the formation of 3 groups of antibody; the group of type-specific antibody was present in a higher titre than the other two antibody groups. isolate 63/70, representing serotype c ... | 1982 | 6188354 |
| serotypes of red clover necrotic mosaic virus. ii. typing of 34 isolates. | examination of 34 isolates of red clover necrotic mosaic virus (rcnmv) from czechoslovakia in agar gel double diffusion precipitation tests and immunoelectrophoresis in agarose gel showed that 16 isolates belonged to serotype b, 6 isolates to serotype c and 4 isolates to serotype a; 3, 1 and 4 isolates represented mixtures of serotypes a + b, a + c and b + c, respectively. the distribution of the individual rcnmv serotypes in czechoslovakia is not bound to definite geographic areas; two serotype ... | 1982 | 6132544 |
| the bipartite genome of red clover necrotic mosaic virus. | purified preparations of red clover necrotic mosaic virus isolated in australia have been shown to contain three rna components whose electrophoretic mobilities in polyacrylamide gel electrophoresis indicate molecular weights of 1.5 x 106 (rna 1), 0.5 x 10(6) (rna 2), and 0.14 x 10(6) (rna 3). comparisons of the rnas by hybridization analysis with 3h-labeled complementary dnas synthesized in vitro have established that rnas 1 and 2 are unique rna molecular species with little or no sequence homo ... | 1981 | 18635032 |
| some properties of red clover necrotic mosaic virus. | | 1969 | 4389117 |