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pathways of degradation of brome mosaic virus particles as influenced by their rna content. 19763877
the structure of a spherical plant virus (bromegrass mosaic virus) established by neutron diffraction.the localization of the rna in a spherical plant virus is established by neutron diffraction. for bmv, this localization is different at low ph than it is at high ph (swollen virus).197613421
swelling of brome mosaic virus as studied by intensity fluctuation spectroscopy. 197720511
[solubilization of a viro-specific rna polymerase activity by treatment with detergents from extracts of barley leaves infected by brome mosaic virus]. 197558633
detection and sequential appearance of typically viral rna species after brome mosaic virus infection in vivo. 197664163
observations of antigenic relatedness between viruses of the herpes simplex "neutroseron".the antigenic relatedness of three viruses of the herpes simplex type 1 neutroseron - herpes simplex virus types 1 (hsv-1) and 2 (hsv-2) and bovine mammillitis virus (bmv) - has been examined by immune precipitation and virus neutralization tests. many virus-specific infected-cell polypeptides were shown to possess antigenic sites shared by both hsv-1 and hsv-2. cross-neutralization between the viruses is mediated through antibodies to at least two antigenic sites, one shared by hsv-1, hsv-2 and ...197887364
blocked 5' termini in brome mosaic virus rna.all four components of brome mosaic virus rna have m(7)g(5') ppp (5')gp as their 5' terminus. the m(7)g can be removed by beta-elimination, resulting in the conversion to pppgp.1976176464
location of the sequences coding for capsid proteins vp1 and vp2 on polyoma virus dna.the 19s and 16s polyoma virus late mrnas have been separated on sucrose-formamide density gradients and translated in vitro. the 16s rna codes only for polyoma capsid protein vp1, while the 19s rna codes in addition for capsid protein vp2. since the 19s and 16s species have been previously mapped on the viral genome, these results allow us to deduce the location of the sequences coding for vp1 and vp2. comparison of the chain lengths of the capsid proteins with the size of the viral mrnas coding ...1976186193
anatomy of bovine mammillitis dna. i restriction endonuclease maps of four populations of molecules that differ in the relative orientation of their long and short components.in this paper, we report that the dna of bovine mammillitis virus (bmv) consists of two covalently linked components that are 71.5 x 10(6) and 15.7 x 10(6) in molecular weight and designated l and s, respectively. we further report that the bmv dna consists of four equimolar populations differing only in the orientation of the l and s components relative to each other. this conclusion is based on the following: (i) the sum molecular weight of fragments generated by digestion of bmv dna with hsu ...1978202750
anatomy of bovine mammillitis dna ii. size and arrangements of the deoxynucleotide sequences.we previously reported that bovine mammillitis virus (bmv) dna consists of two covalently linked components designated l and s and estimated to be 71.5 x 10(6) and 15.7 x 10(6) in molecular weight, respectively; the components invert relative to each other, giving rise to four equimolar populations differing soley in the relative orientation of the two components. we now report that (i) bmv dna has a contour length corresponding to a molecular weight of 89 x 10(6). (ii) component l consists of a ...1978211253
efficient ribosome binding of brome mosaic virus (bmv) rna4 contributes to its ability to outcompete the other bmv rnas for translation.analysis of translation products synthesized in vitro in the presence of a mixture of brome mosaic virus (bmv) rnas 1, 2, 3, and 4 usually shows a predominance of coat protein, coded by rna4. the proportion of products directed by rnas 1 and 2 decreased at saturating concentrations of bmv rna and in the presence of 7-methylguanosine 5'-phosphate (m7g5'p) at nonsaturating concentrations of rna. no differences in the relative proportions of the various proteins produced were detected in the presen ...1979429140
specific inhibition of capped mrna translation in vitro by m7g5'pppp5'g and m7g5'pppp5'm7g.a unique set of diguanosine cap analogues containing a 5'-5' tetraphosphate linkage instead of the normal triphosphate was synthesized by chemical methylation of g5'pppp5'g. both 7-methylguanosine products, m7g5'pppp5'g and m7g5'pppp5'm7g, acted as potent inhibitors of capped brome mosaic virus (bmv) rna translation in the homologous wheat germ protein synthesis system. inhibition of in vitro protein synthesis required the presence of the 7-methyl group on guanosine and was specific for capped m ...1979493116
two-step binding of eukaryotic ribosomes to brome mosaic virus rna3.although brome mosaic virus rna3 has only one translatable cistron, it can bind two 80s ribosomes at initiation. one ribosome binds at the first aug codon (base 92-94). the other binds nearer the 5' end at an entry or holding site. disome formation is thus unrelated to a silent cistron approximately 1,000 bases downstream.1979551276
conformational states of cowpea chlorotic mottle virus ribonucleic acid components.the conditions determining conformational changes in the four ribonucleic acid components of cowpea chlorotic mottle virus have been studied. all four components have at least two electrophoretically separable conformers, the occurrence of which can be regulated by both monovalent and polyvalent cations. this phenomenon also occurs, in a much less striking way, in the ribonucleic acids of the two other members of the bromovirus group, brome mosaic virus and broad bean mottle virus. although spec ...1978643624
in vitro encapsidation of the four rna species of brome mosaic virus. 1978664230
nucleotide sequence of a viral rna fragment that binds to eukaryotic ribosomes.the nucleotide sequence has been determined for the first 53 bases of brome mosaic virus rna4, the monocistronic messenger for brome mosaic virus coat protein. the sequence includes the binding site for wheat embryo ribosomes. the 5'-terminal base is a modified guanosine attached to the penultimate base through a 5' p-p-p 5' link. the initiating aug codon is only 10 nucleotides from the 5'-terminus. the triplets following the aug codon correspond to the known sequence of brome mosaic virus coat ...1975807854
a possible replicative form of brome mosaic virus rna 4. 1976948871
translation of the rnas of brome mosaic virus: the monocistronic nature of rna1 and rna2. 1976957426
interaction of elongation factor 1 with aminoacylated brome mosaic virus and trna's.tyrosylated brome mosaic virus rna was found to interact with a binary complex of wheat germ, elongation factor 1 and [3h]gtp. increasing amounts of the aminoacylated viral rna proportionately reduced radioactivity bound to a nitrocellulose filter, as has previously been noted by others for the charged forms of tobacco mosaic virus, turnip yellow mosaic virus, and trna's. however, sephadex chromatography of the products showed that instead of forming the ternary complex elongation factor-gtp-ami ...1976978788
structure and function of the rnas of brome mosaic virus. 19761019357
translation of virus mrna: comparison of reovirus and brome mosaic virus single-stranded rnas in a wheat germ cell-free system. 19751148014
effect of plumbous ion on messenger rna.the effects of pb2+, a potent catalyst for the depolymerization of rna have been studied on brome mosaic virus (bmv) rna, rabbit globin m-rna and polyuridylic acid. after exposure of these natural and synthetic messengers to a sufficiently high concentration of pb2+, they all lost their ability to stimulate amino acid incorporation in cell-free protein-synthesizing systems. there were differences in the susceptibilities of the messengers; gloing the m-rna for 40 min revealed that there was a thr ...19751175247
evaluation of the techniques used for investigating the in vitro synthesis of bromegrass mosaic virus rna.the paper is dealing with the study of rna polymerase activity using endogenous rna templates in extracts of leaves infected with bromegrass mosaic virus (bmv). the in vitro labeling of bmv-rna in such extracts is associated with a particulate replication complex which is able to synthesize totally or partially single-stranded bmv-rna under conditions where exogenous bmv-rna (purified from the virus) is rapidly degraded. this suggests that rapidly-labeled bmv-rna is protected from endogenous nuc ...19761275388
molecular design of a eukaryotic messenger rna and its chemical synthesis.a designed mrna consisting of 42 ribonucleotides having the cap structure was synthesized. the capped leader sequence of the brome mosaic virus (bmv) mrna 4, m7g5'pppguauuaaua (f-1), was synthesized by the phosphotriester method and followed by the capping reaction. a 32-mer consisting of an initiation codon (aug), the coding region corresponding to a bacterial pheromone cad1 and two stop codons, was constructed by the 18-mer (f-2) and 14-mer (f-3), which were synthesized by the phosphoramidite ...19921374557
exogenous primer-independent cdna synthesis with commercial reverse transcriptase preparations on plant virus rna templates.upon reverse transcription and cloning manipulations with virion rnas of several plant viruses, namely beet yellows virus, brome mosaic virus, and potato virus x, we came across a significant background synthesis of cdna on the virion rna template in vitro independent of exogenous primers added. when tested with beet yellow virus rna template, several commercial preparations of avian myeloblastosis virus (amv) reverse transcriptase showed the background activity monitored by the [alpha-32p]dntp ...19921381874
brome mosaic virus rna replication proteins 1a and 2a from a complex in vitro.brome mosaic virus (bmv) is a positive-strand rna virus that encodes two rna replication proteins, the helicaselike 1a and the polymeraselike 2a. 1a and 2a share extensive sequence similarities with proteins encoded by many other members of the alphaviruslike superfamily. while further purifying enzymatically active rna-dependent rna polymerase from plants infected by bmv, we observed that 1a, 2a, and the polymerase activity all cofractionated through multiple independent purification steps. mor ...19921404594
genetic recombination in brome mosaic virus: effect of sequence and replication of rna on accumulation of recombinants.in order to facilitate the isolation of recombinants in brome mosaic virus, a series of duplication mutants with alterations in the rna3 3' noncoding region has been engineered. the distribution of crossovers, which was observed to be dependent on the parental rna3 sequence, supported the role of rna structure in recombination. however, a negative correlation between replication of the parental rna3 constructs and the accumulation of recombinant progeny confirmed the role of selection.19921404621
the nucleotide sequence of rna-1 of raspberry bushy dwarf virus.raspberry bushy dwarf virus (rbdv) has isometric, 33 nm diameter particles and a bipartite rna genome. sequencing of the larger component (rna-1) showed that it consists of 5449 nucleotides and contains one large open reading frame encoding a putative translation product with a calculated m(r) of 190,000. comparisons of this polypeptide with non-structural proteins of other plant viruses revealed significant homologies with those of alfalfa mosaic virus (almv), brome mosaic virus (bmv), cucumber ...19921469359
the nucleotide sequence and genome organization of the rna2 and rna3 segments in broad bean mottle virus.complete nucleotide sequences of broad bean mottle virus (bbmv) genomic rnas 2 and 3 were determined. they consist of 2811 and 2293 nucleotides, respectively. both rnas are caped and, unlike in other tricornaviruses, both initiate with an a residue. bbmv rna2 is monocistronic and encodes an 815 amino acid 2a protein, whereas rna3 is dicistronic, encoding for a 295 amino acid 3a protein and for the 190 amino acid coat protein. a central, 423 amino acid 2a protein core region is highly homologous ...19921546462
a virus-particle vaccine prepared from bovine mammillitis virus against herpes genitalis.a vaccine against herpes genitalis was prepared from the extracellular virus particles from baby hamster kidney cells infected with bovine mammillitis virus (bmv) strain "allerton". the virus was inactivated by formaldehyde followed by ultracentrifugation to concentrate the virus particles and eliminate formaldehyde to an acceptable concentration for immunisation of human subjects. the vaccine was cross antigenic and cross immunogenic with herpes simplex virus type 1. thirty-four consorts at hig ...19911657508
expression of brome mosaic virus-encoded replicase genes in transgenic tobacco plants.we introduced replicase genes of brome mosaic virus (bmv) to nicotiana tabacum cv. petit habana (sr1) using two different types of transformation vectors containing cdnas of bmv rna 1 and rna 2. one type (v type) contains cdna from which complete viral rnas are transcribed. these rnas can function as templates for viral replicase. the other type (m type) contains cdna from which viral rnas without their 3' non-coding regions are transcribed; these rnas can only function as mrna. viral replicase ...19921730938
the requirement for a 5' stem-loop structure in brome mosaic virus replication supports a new model for viral positive-strand rna initiation.sequences with strong similarity to internal control regions 1 and 2 (icr1 and -2; a and b boxes) of trna genes are found at the 5' termini of the genomic rnas of brome mosaic virus (bmv) and other plant viruses. the functionality of these motifs was studied by introducing point mutations into the icr2-like sequence of prna-2 m/s, a bmv rna-2 deletion mutant that replicates in the presence of rnas-1 and -2 but does not encode a functional viral protein. the accumulation of positive-strand progen ...19921731107
artificial defective interfering rnas derived from brome mosaic virus.naturally occurring defective interfering rnas (di-rnas) greatly reduce the accumulation of their helper virus in vivo, but are rarely associated with plant positive-strand rna viruses. deletion mutants prna-2 m/s and prna-2 e/s, derived from brome mosaic virus (bmv) genomic rna-2, replicated in a manner dependent on bmv rna-1 and -2, and effectively interfered with their accumulation in barley protoplasts. based on their mode of replication, these mutant rnas have been termed parasitic rnas (pr ...19911875191
evolutionary relationships in the cucumoviruses: nucleotide sequence of tomato aspermy virus rna 1.rna 1 of the v strain of tomato aspermy virus (tav) consists of 3410 nucleotides and contains one open reading frame (orf) of 2982 nucleotides, resembling rna 1 of cucumber mosaic virus (cmv) strains q and fny (68% and 66% identical, respectively) and of brome mosaic virus (bmv) (41% identical). in comparisons between amino acid sequences, three conserved regions (n-terminal, c-terminal and central) between tav and each cmv were found. the n- and c-terminal regions were also conserved with bmv, ...19911895056
non-replicating deletion mutants of brome mosaic virus rna-2 interfere with viral replication.naturally occurring defective interfering rnas (di-rnas) and satellite rnas greatly reduce the accumulation of their helper virus in vivo, but often modulate symptom expression in an unpredictable manner. deletion mutants nc/s, na/m and sa/nc + m/s, derived from brome mosaic virus (bmv) rna-2, failed to replicate when co-inoculated with bmv rnas-1 and -2 to barley protoplasts. however, the inoculum rna corresponding to these deletion mutants was extremely stable and could have been mistaken for ...19911919522
tomato aspermy virus has an evolutionary relationship with other tripartite rna plant viruses.the entire rna 3 (2214 nucleotides) of a chrysanthemum isolate of tomato aspermy virus (c-tav) has been cloned and its sequence determined. c-tav possesses two open reading frames which encode a 3a protein (277 amino acids) and a coat protein (229 amino acids). computer-assisted comparisons were made between c-tav rna 3 and its predicted protein sequences and those of two other tripartite rna viruses, cucumber mosaic virus (cmv) and brome mosaic virus (bmv). results from this study suggest that ...19911990057
infectivity of plasmids containing brome mosaic virus cdna linked to the cauliflower mosaic virus 35s rna promoter.full-length biologically active cdnas of brome mosaic virus genomic rnas 1, 2 and 3 were constructed by joining cdna fragments. the cdnas were constructed so that, at the 5' ends, unique snabi sites were present at the site of initiation of transcription. the cdnas were inserted between a modified cauliflower mosaic virus (camv) 35s rna promoter and terminator regions derived from camv dna, and cloned into puc18. when a mixture of the plasmid dnas was inoculated onto chenopodium hybridum leaves, ...19911993867
nucleotide sequence of tomato aspermy virus rna 2.rna 2 of the v strain of tomato aspermy virus (tav) consists of 3074 nucleotides and contains one open reading frame of 2487 nucleotides. thus, it resembles rna 2 of cucumber mosaic virus (cmv) strains q and fny (62% identical to both), brome mosaic virus (42% identical) and cowpea chlorotic mottle virus (40% identical). in comparisons between amino acid sequences, three different regions of similarity could be distinguished. these were the central part (amino acids 224 to 757 for v-tav), which ...19912016592
analysis of the protein composition of alfalfa mosaic virus rna-dependent rna polymerase.rna-dependent rna polymerase (rdrp) was solubilized and purified from cellular membranes isolated from alfalfa mosaic virus (aimv)-infected tobacco by employing a procedure recently described for brome mosaic virus rdrp [r. quadt and e.m.j. jaspars, 1990, virology 178, 189-194]. the purified aimv rdrp is completely dependent on added template rnas and exhibits a high degree of template specificity. analysis of the protein composition of aimv rdrp showed that aimv-encoded proteins p1 and p2 and t ...19912024468
regulation of (+):(-)-strand asymmetry in replication of brome mosaic virus rna.transfection of barley protoplasts with brome mosaic virus (bmv) rnas 1 + 2 in the absence of rna-3 yielded a molar ratio for (+):(-)-strand progeny at 24 hr postinoculation near unity, whereas over 100-fold more (+)- than (-)-strand progeny accumulated in its presence. the presence of rna-3 enhanced total (+)-strand rna production 205-fold and that of rnas 1 + 2 by 29-fold. in contrast, total (-)-strand rna accumulation decreased by 68% and that for (-)rnas 1 + 2 by 79% in the presence of rna-3 ...19912024481
deletion analysis of brome mosaic virus 2a protein: effects on rna replication and systemic spread.brome mosaic virus (bmv) genomic rna2 encodes the 94-kda 2a protein, which is one of two bmv nonstructural proteins required for rna replication and subgenomic mrna transcription. 2a contains a central polymeraselike region, which has extensive sequence similarity with the sindbis virus nsp4 and tobacco mosaic virus (tmv) 183-kda replication proteins, and also contains n- and c-terminal flanking segments without counterparts in the sindbis virus and tmv nonstructural proteins. to further investi ...19912033655
turnip yellow mosaic virus rnas with anticodon loop substitutions that result in decreased valylation fail to replicate efficiently.single and multiple nucleotide substitutions have been introduced into the anticodon loop of the trna-like structure of turnip yellow mosaic virus (tymv) genomic rna. we studied the effects of these mutations on in vitro valylation and on replication in chinese cabbage protoplasts and plants. only those mutants capable of efficient and complete valylation showed efficient replication in protoplasts and gave rise to systemic symptoms in whole plants. mutants that accepted valine inefficiently (in ...19912033666
biological activities of hybrid rnas generated by 3'-end exchanges between tobacco mosaic and brome mosaic viruses.sequences within the conserved, aminoacylatable 3' noncoding regions of brome mosaic virus (bmv) genomic rnas 1, 2, and 3 direct initiation of negative-strand synthesis by bmv polymerase extracts and, like sequences at the structurally divergent but aminoacylatable 3' end of tobacco mosaic virus (tmv) rna, are required in cis for rna replication in vivo. a series of chimeric rnas in which selected 3' segments were exchanged between the tyrosine-accepting bmv and histidine-accepting tmv rnas were ...19912041076
initiation of translation directed by 42s and 26s rnas from semliki forest virus in vitro.the proteins synthesized in vitro in response to 42s and 26s rnas from semliki forest virus were labeled with formyl-[35s]methionine from initiator trna. one protein which comigrated with viral capsid protein was labeled under the direction of 26s rna, and only one labeled peptide was detected after digestion with trypsin. further digestion with pronase gave rise to the dipeptide fmet-asn. several labeled polypeptides were found in the 42s rna directed product and these had molecular weights of ...19761067601
the multipartite genome of brome mosaic virus: aspects of in vitro translation and rna structure.the multipartite nature of the brome mosaic virus genome is described. particular attention is given to the translation of the rnas in cell-free extracts, from dry wheat seed embryos and commercial wheat germ. such studies have demonstrated that the smallest rna is a monocistronic message for coat protein. this cistron is also carried on the intermediate sized rna, which also codes for a larger protein, of unknown function. translation of the larger rnas 1 and 2 is compared, incell-free extracts ...1976776056
frog virus 3-mediated translational shut-off: frog virus 3 messages are translationally more efficient than host and heterologous viral messages under conditions of increased translational stress.frog virus 3 rapidly and selectively blocks host cell translation while synthesizing more than 60 virus-specific polypeptides. previous work indicated that virus infection led to activation of a kinase that phosphorylated and, as a consequence, inactivated eif-2. although phosphorylation of eif-2 could explain the rapid decline in host cell translation, it could not explain how viral protein synthesis persisted in the face of host shut-off. to explain this phenomenon, we speculated that viral me ...19902201134
translational regulation of expression of the brome-mosaic-virus rna genome in vitro.total brome mosaic virus (bmv) rna, comprising four mrna molecules, was translated in the wheat germ system. it was shown that at a low ratio of total bmv rna to cell-free extract all four bmv genes were expressed. at a high excess of total bmv rna over the cell-free extract, the only bmv gene translated was the coat protein cistron (rna 4). the effect was due to diminished initiation of synthesis of non-coat proteins in the presence of a high excess of template over 23 000 x g extract. this lea ...1978658054
the preparation and characterization of a cell-free system from saccharomyces cerevisiae that translates natural messenger ribonucleic acid.a cell-free protein-synthesizing system has been prepared from saccharomyces cerevisiae by differential centrifugation of lysed spheroplasts. the preparation, a modified 100,000 x g supernatant fraction, contains ribosomes and monosomes, ribosomal subunits, translation factors, and aminoacyl-trna synthetases, but no polysomes. after removal of small amounts of remaining mrna with micrococcal nuclease, protein synthesis is stringently dependent on the addition of mrna, as well as amino acids and ...1979374404
homologous nucleotide sequences between prokaryotic and eukaryotic mrnas: the 5'-end sequence of the mrna of the lipoprotein of the escherichia coli outer membrane.the sequence of the first 89 nucleotides at the 5' end of the mrna for the lipoprotein of the escherichia coli outer membrane is: gcuacauggagauuaacucaaucu-agaggguauuaauaaugaaagcuacuaaacugguacu-gggcgcgguaauccuggguucuacucug. the sequence of the first 72 nucleotides was established by direct sequencing methods and was extended to 89 residues on the basis of the known sequences of oligonucleotides obtained from complete digestion of the mrna by ribonuclease t1 or a and the known amino acid sequence ...1978353808
cis-acting sequences required for in vivo amplification of genomic rna3 are organized differently in related bromoviruses.cowpea chlorotic mottle virus (ccmv) is a positive-strand rna virus that infects dicotyledonous plants. the genome comprises three capped rnas: rna1 (3.2 kb), rna2 (2.9 kb), and rna3 (2.1 kb). cis-acting sequences required for amplification in vivo were explored for rna3, which does not contribute trans-acting factors to viral rna replication. using a ccmv cdna expression system, deletions throughout rna3 were constructed and tested for successful replication in barley protoplasts coinoculated w ...19902305551
highly active template-specific rna-dependent rna polymerase from barley leaves infected with brome mosaic virus.the extraction of a template-dependent and template-specific rna-dependent rna polymerase (nucleosidetriphosphate:rna nucleotidyltransferase, ec 2.7.7.6) from a eukaryotic source is described. the enzyme, extracted from barley leaves infected with brome mosaic virus (bmv), is capable of incorporating high levels of radioactivity into trichloroacetic acid-insoluble products. the purification procedure included solubilization with nonionic detergent and precipitation with polyethylene glycol. the ...1979291912
sequence of an oligonucleotide derived from the 3' end of each of the four brome mosaic viral rnas.a 3'-terminal oligonucleotide fragment, 161 bases long, can be obtained from each of the four brome mosaic virus rnas by means of nuclease digestion. like the four intact brome mosaic virus rnas, each fragment accepts tyrosine in a reaction catalyzed by wheat germ aminoacyl-trna synthetase. the complete nucleotide sequence of the rna 4 fragment has been determined by use of standard radiochemical methods. comparative data for the fragments from rnas 1, 2, and 3 show that they have nearly the sam ...1977270723
loss of infectivity of brome mosaic virus rna after chemical modification of the 3' or 5' terminus.brome mosaic virus (bmv) rna that had both termini chemically modified by periodate oxidation and aniline-catalyzed cleavage of the terminal nucleotide had drastically reduced infectivity. bmv rna that was first enzymatically tyrosylated to protect the 3' terminus from modification, and then modified at the 5' terminus by periodate oxidation and aniline cleavage, had a similar reduction in infectivity. tyrosylation followed by acetylation modifies only the 3' terminus. nevertheless, acetylated t ...1977268617
the effect of conformational changes in brome mosaic virus upon its sensitivity to trypsin, chymotrypsin and ribonuclease. 1975239862
inhibition of rous sarcoma viral rna translation by a specific oligodeoxyribonucleotide.a tridecamer oligodeoxynucleotide, d(a-a-t-g-g-t-a-a-a-a-t-g-g), which is complementary to reiterated 3'- and 5'-terminal nucleotides of rous sarcoma virus 35s rna, is an efficient inhibitor of the translation of proteins specified by the viral rna in the wheat embryo cell-free system. the inhibition specificity for oncornavirus rna is greater than for rabbit reticulocyte mrna or brome mosaic virus rna. other oligodeoxynucleotides of similar size have little or no specific effect on the rna-dire ...197875546
use of bromovirus rna3 hybrids to study template specificity in viral rna amplification.brome mosaic virus (bmv) and cowpea chlorotic mottle virus (ccmv) are related positive-strand rna viruses with genomes divided among rnas 1, 2, and 3. rnas 1 and 2 encode the viral rna replication factors, which share extensive conservation with proteins encoded by the animal alphaviruses and diverse plant viruses. in barley protoplasts, ccmv rnas 1 and 2 support high but distinguishable amplification of either bmv rna3 (b3) or ccmv rna3 (c3), while bmv rnas 1 and 2 show even greater discriminat ...19912041089
unique nucleotide differences in the conserved 3' termini of brome mosaic virus rnas are maintained through their optimization of genome replication.to explore the functionality and conservation of specific base differences in the 3' 200 nucleotides of brome mosaic virus (bmv) rna-1 (1t) and rna-2 (2t) with respect to the 3' end of rna-3 (3t), all possible permutations were used to exchange these regions among the genomic rnas. when all rnas bore the 1t promoter, total rna accumulation was only 15% of wild type; when the 2t or 3t promoter was present on all three rnas total rna accumulation was reduced to 30 or 35% of wild type. two major pr ...19921736528
requirement for icr-like sequences in the replication of brome mosaic virus genomic rna.previous studies using a brome mosaic virus (bmv) rna-2 deletion mutant (prna-2 m/s) and additional derivatives as reporters established that viral sequences resembling internal control regions (icrs) 1 and 2 of trna gene promoters are vital to (+)-strand replication in protoplasts. transfer of these mutations to genomic rna-2 and functional analysis in protoplast, local lesion, and systemic infections revealed a sequence-specific requirement for bases within the icr2-like motif. despite the low ...19921585645
radioactive labelling of borme mosaic virus.brome mosaic virus (bmv) can be radioactively labelled by allowing virus synthesis to take place in barley leaves whose cut ends are placed in solutions of radio active precursors of bmv. when barley plants are infected by inoculation of the first emerging leaf, the rate of incorporation of both [14c]-leucine and [32p]-ortho-phosphate into virus particles was found to be maximal in the second leaf, cut and labelled 4 to 5 days after inoculation. the rate of incorporation declined when the labell ...19751120938
nucleotide sequence and translation of satellite tobacco mosaic virus rna.satellite tobacco mosaic virus (stmv) is a plant virus with a 17-nm icosahedral particle encapsidating a 0.3 x 10(6) mr ssrna genome that depends on tobamoviruses for its replication. the complete nucleotide sequence of stmv rna deduced in the experiments described here was 1059 nucleotides in length. the efficiency of labeling viral rna with [gamma-32p]atp using t4 polynucleotide kinase was not affected by treatment with tobacco acid pyrophosphatase and/or bacterial alkaline phosphatase, indica ...19892718378
natural fluorescence properties of brome mosaic virus protein.some of the fluorescence properties of brome mosaic virus protein in different states of aggregation (dimer, capsid) have been studied, in particular the emission spectra, fluorescence quantum yields and lifetimes, as well as the effect of external quenchers and of temperature on the fluorescence. brome mosaic virus protein, which contains two tryptophan and five tyrosine residues per monomer, displayed an unusual fluorescence spectrum maximum of 308 nm at ph 7.4 when excited at 280 nm. the emis ...1977880311
telomeric function of the trna-like structure of brome mosaic virus rna.four mutant brome mosaic virus (bmv) rna3 transcripts, bearing single or double base changes in the 3'-ccaoh terminus of the trna-like structure, previously characterized as being deficient in vitro with respect to aminoacylation and replication activities, have now been assayed in vivo for their ability to replicate (in the presence of transcripts of wild-type rna1 and -2) in barley protoplasts and plants. in tests conducted with protoplasts, irrespective of the time post-infection, all four mu ...19892748589
sequence of cowpea chlorotic mottle virus rnas 2 and 3 and evidence of a recombination event during bromovirus evolution.the genomic sequence of cowpea chlorotic mottle virus (ccmv) was completed by sequencing biologically active cdna clones of ccmv rna2 (2774 bases) and rna3 (2173 bases). while only the central core of the encoded 94-kda ccmv 2a protein contains features conserved among known and putative rna replication proteins from many viruses, both flanking regions of ccmv 2a show substantial similarity to the corresponding protein of the related brome mosaic virus (bmv). the 3a proteins of ccmv and bmv, imp ...19892773323
mrna(nucleoside-2'-)-methyltransferase from vaccinia virus. characteristics and substrate specificity.an mrna(nucleoside-2'-)-methyltransferase purified from vaccinia virus was shown to methylate the penultimate nucleoside of rna ending in m7g(5')pppn-. by contrast, rnas ending in pn-, ppn-, or even g(5')pppn- are not methyl acceptors. this specificity indicates that 2'-o-methylation is the final step in the formation of the m7g(5')pppnm- cap structure. both adenosine and guanosine are methylated, in accordance with the presence of these nucleosides in the penultimate position of vaccinia virus ...1978701282
mrna(nucleoside-2'-)-methyltransferase from vaccinia virus. purification and physical properties.an s-adenosyl-l-methionine:mrna(nucleoside-2'-)-methyltransferase, one of at least three activities required for the 5'-terminal modification of mrna, has been purified from vaccinia virus particles. employing brome mosaic virus rna ending in m7g(5')pppg- as substrate, a simple deae-cellulose filter assay measuring the incorporation of methyl groups from s-adenosyl[methyl-3h]methionine to position 2' of the penultimate nucleoside was devised. starting from disrupted vaccinia virus cores, a 350-f ...1978701281
ultrasonic absorption evidence of structural fluctuations in viral capsids.when the coat protein of the small icosahedral virus, brome mosaic virus, reassembles into capsids, the ultrasonic absorption of the solution greatly increases. submitting the solution to an ultrasonic field thus appears to reveal spontaneous molecular motions within a protein assembly. confirmatory evidence of a dynamics of a protein shell comes from measurements on brome mosaic virus at various degrees of swelling and on tomato bushy stunt virus treated with the crosslinking agent glutaraldehy ...1979287017
studies on the mechanism of translational enhancement by the 5'-leader sequence of tobacco mosaic virus rna.translation of foreign mrnas is enhanced by a cis-acting derivative (omega') of the 5'-leader sequence (omega) of tobacco mosaic virus rna (vulgare strain). to explain this effect we have conducted several experiments in vitro. 1. the presence of various 5'-terminal sequences, including omega', did not significantly increase the half-lives of chloramphenicol acetyltransferase (cat) or neomycin phosphotransferase (nptii) mrnas in wheat-germ extract. also, a long leader sequence, unrelated to omeg ...19882841127
structural analogies between the 3' trna-like structure of brome mosaic virus rna and yeast trnatyr revealed by protection studies with yeast tyrosyl-trna synthetase.contacts between the trna-like domain in brome mosaic virus rna and yeast tyrosyl-trna synthetase have been determined by footprinting with enzymatic probes. regions in which the synthetase caused protections indicative of direct interaction coincide with loci identified by mutational studies as being important for efficient tyrosylation [dreher, t. w. & hall, t. c. (1988) j. mol. biol. 201, 41-55]. additional extensive contacts were found upstream of the core of the trna-like structure. in para ...19892684668
reduced tumorigenicity of rodent tumour cells and tumour explants following infection with wild type and mutant herpes simplex virus, bovine mammillitis virus and encephalomyocarditis virus.the tumorigenicity of neoplastic hamster and mouse cell lines and tumour explants was reduced by infection with herpes simplex virus (hsv-1), a thymidine-kinaseless mutant of herpes simplex virus, namely 'mdk', encephalomyocarditis virus (emc) and bovine mammillitis virus (bmv). there was an approximate relationship between duration of virus infection in vitro and reduction in incidence and/or rate of tumour development. the rate of tumour development was also reduced by 'site inoculation' of vi ...19882846027
hybrid brome mosaic virus rnas express and are packaged in tobacco mosaic virus coat protein in vivo.brome mosaic virus (bmv) is an icosahedral virus with a tripartite rna genome which infects monocotyledonous plants, while the cowpea or legume strain of tobacco mosaic virus (cctmv) is a rod-shaped virus with a single component rna genome which infects dicotyledonous plants. to examine the potential for exchanging entire genes between rna viruses, biologically active cdna clones were used to replace the natural coat gene of bmv rna3 with the coat gene and encapsidation origin of cctmv. in proto ...19882847411
analysis of rna stability and (-) strand content in viral infections using biotinylated rna probes.non-radioactive biotinylated rna probes specific for plus (+) and minus (-) sense rnas of brome mosaic virus (bmv) were synthesized in vitro from a plasmid bearing a 200 base pair fragment complementary to the 3' terminus of each of the three genomic rnas of the virus. using virion rnas isolated from bmv infected barley plants, the sensitivity of biotinylated rnas as hybridization probes was compared with that of 32p-labeled probes in northern hybridization assays. although the sensitivity of bi ...19901707891
a neurospora crassa heat-shocked cell lysate translates homologous and heterologous messenger rna efficiently, without preference for heat shock messages.cell-free protein synthesis systems were prepared from normally-grown (n-lysate) and heat-shocked (hs-lysate) neurospora crassa mycelium. although both lysates translated homologous mrna, the hs-lysate was more active, yielding a higher incorporation of [35s]-methionine into hot tca-insoluble material and a vastly superior protein synthesis profile. the optimal temperature for translation by both lysates was 21 degrees c; the hs-lysate did not translate heat-shock mrna preferentially at any temp ...19882969781
antigenic and biochemical analysis of gb of herpes simplex virus type 1 and type 2 and of cross-reacting glycoproteins induced by bovine mammillitis virus and equine herpesvirus type 1.an antiserum was produced to the oligomeric form of glycoprotein b (gb) induced by herpes simplex virus type 1 (hsv-1) strain 17. this antiserum gave a single common precipitin line in agar gel immunodiffusion with hsv-1, hsv-2, bovine mammillitis virus (bmv) and equine herpesvirus type 1 (ehv-1). it also neutralized hsv-1, hsv-2 and bmv but not ehv-1. absorption of the antiserum with excess hsv-2 or bmv antigen resulted in an hsv-1-specific neutralizing antiserum. in immunoprecipitation, two pr ...19852981966
role of bovine mammillitis virus towards preparation of an alternative vaccine against herpes simplex virus infections of human subjects.bovine mammillitis virus (bmv) cross-reacted in neutralization and radioimmune assay with herpes simplex virus (hsv) and pre-immunization with bmv protected against challenge by type 2 hsv. there was no evidence to suggest a pathogenic role for bmv as adjudged by a literature search or field enquiry and bmv specific antibody was not detected in 21 human sera or in four sera from personnel engaged in research with bmv; in addition there was no replication or antigen synthesis by bmv in explants o ...19873033927
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 ...19883047682
a continuous cell-free translation system capable of producing polypeptides in high yield.a cell-free translation system has been constructed that uses a continuous flow of the feeding buffer [including amino acids, adenosine triphosphate (atp), and guanosine triphosphate (gtp)] through the reaction mixture and a continuous removal of a polypeptide product. both prokaryotic (escherichia coli) and eukaryotic (wheat embryos, triticum sp.) versions of the system have been tested. in both cases the system has proven active for long times, synthesizing polypeptides at a high constant rate ...19883055301
hemolysates from guinea-pig reticulocytes also efficiently translate added mrna.1. lysate is prepared from guinea-pig reticulocytes and its translation activity is compared both quantitatively and qualitatively with that found in rabbit reticulocyte lysates. 2. guinea-pigs are comparatively resistant to acetylphenylhydrazine, a drug used to induce reticulocytosis in rabbits. 3. the characteristics and conditions for maximal translation activity in guinea-pig reticulocyte lysates are quite similar to those described for rabbit reticulocyte systems with the possible exception ...19883197393
nucleotide sequence responsible for the synthesis of a truncated coat protein of brome mosaic virus strain atcc66.the atcc66 strain of brome mosaic virus (bmv) (propagated at kyoto university) contained two types of coat protein in its virion whereas the russian strain of bmv has been known to contain a single coat protein; both strains have two initiation codons for coat protein in the same reading frame at the 5'-proximal end of the gene in rna 4. comparative studies on the nucleotide sequences of the atcc66 and russian strains of bmv demonstrated that in the atcc66 strain, two adjacent adenine residues w ...19921402800
hemin and cyclic amp stimulate message-dependent translation in lysates from friend erythroleukemia cells.a message-dependent, cell-free translation system was prepared from both uninduced and n,n'-hexamethylene-bis-acetamide-induced friend erythroleukemia cells following modification of standard protocols. active extracts were prepared by lysing friend erythroleukemia cells in hypotonic buffer containing 50 microm hemin and 10 mm cyclic adenosine monophosphate (camp), and assaying translation in vitro in the absence of exogenous nucleoside triphosphates. both hemin and camp were required for full a ...19892541008
interference in trans with brome mosaic virus replication by rna-2 bearing aminoacylation-deficient mutants.the trna-like domain present at the 3' end of each of the three genomic rnas of brome mosaic virus (bmv) encompasses the (-)-strand promoter essential for replication. the replicative competence of two bmv rna-2 transcripts bearing mutations delta 5' and 5'aga in the trna-like domain (previously shown by in vitro assays to be deficient in tyrosylation) was evaluated in barley protoplasts. transfection of protoplasts with low (2 micrograms) amounts of delta 5'rna-2, together with transcripts of w ...19911984646
complementary oligodeoxynucleotide mediated inhibition of tobacco mosaic virus rna translation in vitro.two different "antisense" oligodeoxynucleotides and their rna analogues, each complementary to non-overlapping sequences of 51 bases near the 5' end of tmv rna, inhibit in vitro translation of the genomic rna in a rabbit reticulocyte lysate. inhibition is dependent upon complementarity, concentration, and hybridization of the oligomers with tmv rna. inhibition is observed at molar ratios of tmv rna to antisense oligomers as low as 1:1.5. a plateau of inhibition at which 10-25% of the control sig ...19883380688
further implications for the evolutionary relationships between tripartite plant viruses based on cucumber mosaic virus rna 3.the nucleotide sequence of the rna 3 of the q-strain of cucumber mosaic virus (q-cmv) has been reinvestigated and supporting partial amino acid sequence data obtained for the coat protein. corrections to the previously published sequence of rna 3 [a. r. gould and r. h. symons (1982) eur. j. biochem. 126, 217-226] result in changes to the size and composition of the putative 3a and coat proteins. analysis of the nucleotide sequence revealed a 14-nucleotide sequence present in the intercistronic r ...19883388769
involvement of a nonstructural protein in the rna synthesis of brome mosaic virus.rna-dependent rna polymerase (rdrp) was prepared from brome mosaic virus (bmv)-infected barley by a procedure including nonidet-p40 treatment. the enzyme proved to be highly active, specific, and almost completely template dependent without the need for nuclease treatment [w. a. miller, and t. c. hall (1983) virology 125, 236-241] or deae ion exchange chromatography [k. maekawa and i. furusawa (1984) ann. phytopathol. soc. japan 50, 491-499]. two c-terminal peptides p1c and p2c derived from the ...19883388771
the interaction of wheat germ tyrosyl-trna synthetase and the trna-like end of brome mosaic virus rna has no effect on in vitro viral protein synthesis and on in vitro encapsidation.the effect of aminoacylation of the trna-like end of brome mosaic virus rna during in vitro protein synthesis and in vitro viral encapsidation was investigated. the components of the homologous system were: bmv rna, wheat germ cell-free protein synthesizing system and pure tyrosyl-trna synthetase from wheat germ. during in vitro protein synthesis directed with tyrosylated as well as non-tyrosylated bmv rna, no differences were observed in the amount and in the class of polypeptides formed neithe ...19911782218
molecular studies of brome mosaic virus using infectious transcripts from cloned cdna. 19873303861
effects of deletions in the n-terminal basic arm of brome mosaic virus coat protein on rna packaging and systemic infection.the first 25 amino acids of brome mosaic virus (bmv) coat protein include 8 basic and no acidic residues and are implicated in binding the encapsidated rna. using infectious transcripts from bmv rna3 cdna clones, we modified this region of the coat gene. a coat protein mutant with the first 25 amino acids deleted failed to direct either packaging of viral rna in protoplasts or systemic infection of whole barley plants. neither symptoms, virions, nor viral rna was detectable in plants inoculated ...19892795712
search of essential parameters for the aminoacylation of viral trna-like molecules. comparison with canonical transfer rnas.comparative structural and functional results on the valine and tyrosine accepting trna-like molecules from turnip yellow mosaic virus (tymv) and brome mosaic virus (bmv), and the corresponding cognate yeast trnas are presented. novel experiments on tymv rna include design of variant genes of the trna-like domain and their transcription in vitro by t7 rna polymerase, analysis of their valylation catalyzed by yeast valyl-trna synthetase, and structural mapping with dimethyl sulfate and carbodiimi ...19902207141
intercistronic as well as terminal sequences are required for efficient amplification of brome mosaic virus rna3.the genome of brome mosaic virus (bmv) is divided among messenger polarity rna1, rna2, and rna3 (3.2, 2.9, and 2.1 kilobases, respectively). cis-acting sequences required for bmv rna amplification were investigated with rna3. by using expressible cdna clones, deletions were constructed throughout rna3 and tested in barley protoplasts coinoculated with rna1 and rna2. in contrast to requirements for 5'- and 3'-terminal noncoding sequences, either of the two rna3 coding regions can be deleted indiv ...19873573144
the common 5' terminal sequence on trypanosome mrnas: a target for anti-messenger oligodeoxynucleotides.several mature mrnas of trypanosoma brucei were previously shown to have a common 5' terminal sequence of 35 nucleotides (nt) encoded by a separate mini-exon. to verify whether all trypanosome mrnas contain this mini-exon sequence at their 5' end, we have tested oligodeoxynucleotides complementary to different parts of the 35 nt leader sequence for their ability to inhibit translation of total trypanosome mrna. all oligomers tested inhibited translation of trypanosome mrnas in a wheat germ extra ...19863737413
rna viruses: stabilization of brome mosaic virus.the relative importances of protein-protein and rna-protein interactions in stabilizing the architecture of brome mosaic virus particles are discussed in the light of the following experimental evidence: (a) disassembly pathways of the virus particles, (b) reassembly of the virus and self-association capacity of the protein moiety, and (c) the role of divalent cations in virus stabilization, and their relevance to localization of the rna in the virus particles. evidence is given that the capsid ...197613438
identification of the domains required for direct interaction of the helicase-like and polymerase-like rna replication proteins of brome mosaic virus.brome mosaic virus is a positive-strand rna virus whose rna replication requires viral protein 1a, which has putative helicase and capping functions, and 2a, which has putative polymerase function. since domains of related sequence are conserved in a wide range of plus-strand rna viruses, analysis of 1a and 2a function should have applicability to many other viruses. we have recently demonstrated that 1a and 2a form a complex in vivo and in vitro. using immune coprecipitation and mutant polypept ...19921433519
generation and analysis of nonhomologous rna-rna recombinants in brome mosaic virus: sequence complementarities at crossover sites.all three single-stranded rnas of the brome mosaic virus (bmv) genome contain a highly conserved, 193-base 3' noncoding region. to study the recombination between individual bmv rna components, barley plants were infected with a mixture of in vitro-transcribed wild-type bmv rnas 1 and 2 and an rna3 mutant that carried a deletion near the 3' end. this generated a population of both homologous and nonhomologous 3' recombinant bmv rna3 variants. sequencing revealed that these recombinants were deri ...19912072447
analysis of the role of brome mosaic virus 1a protein domains in rna replication, using linker insertion mutagenesis.brome mosaic virus (bmv) belongs to a "superfamily" of plant and animal positive-strand rna viruses that share, among other features, three large domains of conserved sequence in nonstructural proteins involved in rna replication. two of these domains reside in the 109-kda bmv 1a protein. to examine the role of 1a, we used biologically active cdna clones of bmv rna1 to construct a series of linker insertion mutants bearing two-codon insertions dispersed throughout the 1a gene. the majority of th ...19902243389
the replication of plant virus rna.the roles of plant-encoded rna-dependent rna polymerases and viral rna replicases in the replication of plant viral rna are reviewed with respect to the positive single-stranded tymv, cpmv and bmv.19853940008
use of bromovirus rna2 hybrids to map cis- and trans-acting functions in a conserved rna replication gene.brome mosaic virus (bmv) and cowpea chlorotic mottle virus (ccmv) are related positive-strand rna viruses with tripartite genomes. rna replication by either virus requires genomic rnas 1 and 2, which encode protein 1a and the polymeraselike, 94-kilodalton 2a protein, respectively. proteins 1a and 2a share extensive sequence similarity with proteins encoded by a wide range of other positive-strand rna viruses of animals and plants. heterologous combinations of bmv and ccmv rnas 1 and 2 do not sup ...19902293671
requirement for a viral trans-acting factor encoded by brome mosaic virus rna-2 provides strong selection in vivo for functional recombinants.interaction of specific nucleotide sequences with trans-acting proteins is intrinsic to replication of viral as well as eucaryotic genomes. brome mosaic virus rna-2 encodes one of the two viral proteins known to be essential for replication. (r. french, m. janda, and p. ahlquist, science 231:1294-1297, 1986; p. a. kiberstis, l. s. loesch-fries, and t. c. hall, virology 112:804-808, 1981). transfection of barley protoplasts with wild-type transcripts of brome mosaic virus rna-1 and rna-3 and seri ...19902325208
effect of dinucleotides on wheat germ translation system.the effect of ribodinucleoside monophosphates on total protein synthesis was studied in a wheat germ cell-free system, using brome mosaic virus (bmv) rna as a messenger. dinucleotides inhibit total protein synthesis to different extents. of those tested the most inhibitory is cpa. the inhibitory effect of dinucleotides is due to their adverse effect on initiation and not on elongation of polypeptide synthesis. it seems that the dinucleotides complementary to the initiation codon are able to comp ...19853987903
incomplete polypeptides are formed in vitro by premature chain termination.the mechanism of incomplete polypeptides formation during protein synthesis was studied in the wheat germ cell-free system programmed with brome mosaic virus rna 4. the synthesis of coat protein, the complete product of rna 4 translation, was accompanied by the appearance of polypeptides of lower molecular mass. it was shown that incomplete products are formed by translation of different lengths of rna 4, always from the first 5' aug codon, and were due neither to proteolysis of coat protein nor ...19854006942
nucleotide sequence of cucumber mosaic virus rna. 1. presence of a sequence complementary to part of the viral satellite rna and homologies with other viral rnas.the nucleotide sequence of the 3389 residues of rna 1 (mr 1.15 x 10(6) of the q strain of cucumber mosaic virus (cmv) was determined, completing the primary structure of the cmv genome (8617 nucleotides). cmv rna 1 was sequenced by the dideoxy-chain-termination method using m13 clones carrying rna 1 sequences as well as synthetic oligonucleotide primers on rna 1 as a template. at the 5' end of the rna there are 97 noncoding residues between the cap structure and the first aug (98-100), which is ...19854018086
formation of artificial top component from brome mosaic virus at high salt concentrations. 19744132238
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