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

Morris-Krsinich, Bret A.M.; Forster, R. L. S.; Mossop, D.W.

doi:10.1016/0042-6822(83)90351-3

Cited by 32 publications

(12 citation statements)

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“…wt. 38 000 to 41000, encoded by RNA 1 (Hollings & Stone, 1977;Dodds et al, 1977; Gould et al, 1981 ;Okuno et al, 1983;Morris-Krsinich et al, 1983). The properties of pseudorecombinants formed between RCNMV and two other dianthoviruses, sweet clover necrotic mosaic virus and clover primary leaf necrosis virus, indicated that symptom type in several plant species was determined by RNA 1 or by interaction between RNA 1 and RNA 2 (Okuno et al, 1983).…”

Section: Discussionmentioning

confidence: 99%

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RNA 2 of Red Clover Necrotic Mosaic Virus Determines Lesion Morphology and Systemic Invasion in Cowpea

Osman

Dodd

Buck

1986

Journal of General Virology

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SUMMARYRed clover necrotic mosaic virus (RCNMV) has two RNA species of mol. wt. about 1.5 x 106 (RNA 1) and 0.5 × 106 (RNA 2). An English strain (H) and a Czechoslovakian strain (TpM-34)of RCNMV could be distinguished serologically, by solid-phase RNA hybridization analysis (Northern blotting) and by the symptoms they induced in cowpea. Studies of pseudorecombinants, formed following inoculation of plants with heterologous combinations of the RNA species of each strain, showed that RNA 2 determines the morphology of lesions induced by the isolates in cowpea and their ability to invade the plants systemically.Red clover necrotic mosaic virus (RCNMV) belongs to the dianthovirus group (Matthews, 1982), members of which are characterized by isometric particles 27 to 35 nm in diameter, a genome composed of two single-stranded RNA molecules, RNA 1 (mol. wt. 1.35 x 106 to 1.55 X 106) and RNA 2 (mol. wt. 0.5 × 106 to 0.6 x 106), both of which are required for infectivity, and a single capsid polypeptide species, mol. wt. 38 000 to 41000, encoded by RNA 1 (Hollings & Stone, 1977;Dodds et al., 1977; Gould et al., 1981 ;Okuno et al., 1983;Morris-Krsinich et al., 1983). The properties of pseudorecombinants formed between RCNMV and two other dianthoviruses, sweet clover necrotic mosaic virus and clover primary leaf necrosis virus, indicated that symptom type in several plant species was determined by RNA 1 or by interaction between RNA 1 and RNA 2 (Okuno et al., 1983). We now describe two properties of RCNMV which are determined primarily by RNA 2.English isolate H and Czechoslovakian isolate TpM-34 of RCNMV (Musil, 1969;Hollings & Stone, 1977;Musil & Gallo, 1982) (kindly provided by Drs A. A. Brunt and O. M. Stone) were each subjected to three cycles of single lesion isolation in Chenopodium quinoa to obtain strains as homogeneous as possible. The viruses were propagated in Phaseolus vulgaris (French bean) cv.'The Prince' (Hollings & Stone, 1977) and extracted and purified by differential and sucrose density gradient centrifugations (Gould et al., 1981). Double immunodiffusion analysis of virus particles in 1 ~o agarose gels was as described by Buck et al. (1981). Rabbit antisera to the two virus strains were kindly provided by Dr A. A. Brunt. Capsid polypeptide species were analysed by SDS-PAGE (Laemmli, 1970). RNA was isolated from virus particles by phenol/SDS extraction (Okuno et al., 1983) and the two species were separated by two cycles of centrifugation in sucrose/formamide gradients. Virus RNA in buffered 85 ~ formamide (85 vol. of deionized formamide mixed with 15 vol. 10 mM-Tris-HC1, 1 mM-EDTA, pH 7-5) was heatdenatured (60 °C, 3 min), then cooled to 20 °C and layered onto a 5 ml linear gradient of 2 to 10~ (w/v) sucrose in buffered 85~o formamide. Gradients were centrifuged in a Beckman SW50.1 rotor at 40000 r.p.m, for 17 h at 20°C and then 0.1 ml fractions were collected from the bottom of the tubes. RNA in each fraction was recovered by precipitation with 2.5 vol. ethanol and analysed, after glyoxalation, by el...

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Section: Discussionmentioning

confidence: 99%

“…wt. 34000 (Morris-Krsinich et al, 1983). If most of their sequence were used for coding, RNAs 1 and 2 could encode proteins with total mol.…”

mentioning

confidence: 99%

RNA 2 of Red Clover Necrotic Mosaic Virus Determines Lesion Morphology and Systemic Invasion in Cowpea

Osman

Dodd

Buck

1986

Journal of General Virology

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“…The 57K protein has amino acid sequence motifs characteristic of nucleic acid polymerases (Argos, 1988;Xiong & Lommel, 1989) and helicases (Habili & Symons, 1989;Buck, 1990), suggesting a role in RNA replication; it may be expressed as an 88K fusion protein resulting from a ribosomal frameshift near the end of the ORF for the 27K protein (Xiong & Lommel, 1989). The capsid protein (Mr 37K to 40K) is translated from a subgenomic RNA derived from RNA 1 (Morris-Krsinich et al, 1983;Marriott & Buck, 1988;Osman & Buck, 1990). The proteins encoded by RNA 1 are consistent with its ability to replicate and produce virus particles in plant protoplasts in the absence of RNA 2 (Osman & Buck, 1987;Paje-Manalo & Lommel, 1989).…”

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confidence: 89%

Detection of the movement protein of red clover necrotic mosaic virus in a cell wall fraction from infected Nicotiana clevelandii plants

Osman

Buck

1991

Journal of General Virology

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“…Leaves were ground in 0.5M sodium phosphate buffer, pH 7.1, containing 10 mM ethylenediaminetetraacetic acid (EDT A) and 0.1 % thioglycollic acid (2 ml/g leaves) and chloroform (0.25 ml/g leaves). The extract was centrifuged at 10 OOOg for 10 min and the virus concentrated from the supernatant by centrifugation at 78 OOOg for 3 h. Pellets were resuspended in 1% sodium phosphate (pH 7.1) and the virus further purified on sucrose gradients as described by Morris-Krsinich et al (1983). Serology Antisera to LAL Y and L TSY were produced as described by and .…”

Section: Introductionmentioning

confidence: 99%

Incidence of alfalfa mosaic virus, lucerne Australian latent virus, and lucerne transient streak virus in lucerne crops in the north island of new zealand

Forster

Morris-Krsinich

Musgrave³

1985

New Zealand Journal of Agricultural Research

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The incidence of alfalfa mosaic VIruS (AMY) in lucerne (Medicago sativa L.) Crops in the North Island of New Zealand increased significantly between 1976 and 1982. In contrast lucerne Australian latent virus and lucerne transient streak virus both remained at a low level. Five commercial seed lots of lucerne imported from the United States contained up to 4% seed-borne AMY. Six isolates of AMY from lucerne could be distinguished by symptoms in 5 hosts and by the size and relative amount of in vitro translation products.

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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

Cited by 32 publications

References 14 publications

RNA 2 of Red Clover Necrotic Mosaic Virus Determines Lesion Morphology and Systemic Invasion in Cowpea

RNA 2 of Red Clover Necrotic Mosaic Virus Determines Lesion Morphology and Systemic Invasion in Cowpea

Detection of the movement protein of red clover necrotic mosaic virus in a cell wall fraction from infected Nicotiana clevelandii plants

Incidence of alfalfa mosaic virus, lucerne Australian latent virus, and lucerne transient streak virus in lucerne crops in the north island of new zealand

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