Subcellular distribution of RNA sequences complementary to sonchus yellow net virus RNA

Milner, Joel J.; Hakkaart, Marcel J. J.; Jackson, Andrew O.

doi:10.1016/0042-6822(79)90577-4

Cited by 9 publications

(6 citation statements)

References 12 publications

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“…The present in situ experiments show that both the genomic and the antigenomic RNAs are associated with the nuclei, and they most likely are present as replicating complexes within the viroplasms. The results also complement previous studies showing that the mRNAs accumulate to high abundance in the cytoplasm (30,31). Interestingly, sequences corresponding to the positive strand (antigenomic) leader RNA transcript appear in the cytoplasm as well as in the nucleus.…”

Section: Discussionsupporting

confidence: 90%

“…1 and data not shown). These observations provide additional verification of previous hybridization results indicating cytoplasmic partitioning of polyadenylated SYNV mRNAs (30,31). In addition, our results show unexpectedly that a portion of the polyadenylated leader RNA is present in the cytoplasm (Fig.…”

Section: Genomic and Antigenomic Rnas Are Abundant In Nuclei Of Synv-supporting

confidence: 92%

“…Our studies support a model predicting that early in SYNV infection, virions or viral cores move into the nucleus to estab-lish a viroplasm that functions to mediate transcription of mRNAs as well as the antigenomic and genomic replicative RNAs. According to this model, polyadenylated mRNA transcripts are exported to the cytoplasm, where the mRNAs are translated differentially on free or membrane-bound polysomes (30,38). The present in situ experiments show that both the genomic and the antigenomic RNAs are associated with the nuclei, and they most likely are present as replicating complexes within the viroplasms.…”

Section: Discussionmentioning

confidence: 75%

“…To mediate these integral events in the viral life cycle, the host macromolecular transport machinery must carry out several essential steps requiring nucleocytoplasmic transport of viral components. For example, unspliced SYNV mRNAs are thought to be transcribed in the nucleus (44,45) and exported to the cytoplasm, where they are partitioned into free and membrane-associated polysomes during translation (30,31). After protein synthesis, some of the virus-encoded proteins are postulated to enter the nucleus and participate in formation of the viroplasm complex.…”

mentioning

confidence: 99%

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Sonchus Yellow Net Rhabdovirus Nuclear Viroplasms Contain Polymerase-Associated Proteins

Martins

Johnson

Lawrence

et al. 1998

J Virol

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We have initiated a study of the cytopathology of nucleorhabdoviruses by analyzing the subcellular localization of sonchus yellow net virus (SYNV) genomic and antigenomic RNAs and the encoded polymerase proteins. In situ hybridizations demonstrated that the minus-strand genomic RNA sequences are restricted to the nuclei of infected cells, while the complementary plus-strand antigenomic RNA sequences are present in both the nuclei and the cytoplasm. Immunofluorescence and immunogold labeling experiments also revealed that the nucleocapsid (N) protein and phosphoprotein (M2) are primarily localized to discrete regions within the nuclei and in virus particles that accumulate in perinuclear spaces. The N protein antiserum specifically labeled the nuclear viroplasms, whereas the M2 antiserum was more generally distributed throughout the nuclei. Antibody detection also indicated that the polymerase (L) protein is present in small amounts in the viroplasm. When the N and M2 proteins were expressed individually from the heterologous potato virus X (PVX) vector, both proteins preferentially accumulated in the nuclei. In addition, viroplasm-like inclusions formed in the nuclei of cells infected with the PVX vector containing the N gene. Fusions of the carboxy terminus of β-glucuronidase to N and M2 resulted in staining of the nuclei of infected cells following expression from the PVX vector. Deletion analyses suggested that multiple regions of the N protein contain signals that are important for nuclear localization.

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Section: Discussionsupporting

confidence: 90%

Section: Genomic and Antigenomic Rnas Are Abundant In Nuclei Of Synv-supporting

confidence: 92%

Section: Discussionmentioning

confidence: 75%

mentioning

confidence: 99%

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Sonchus Yellow Net Rhabdovirus Nuclear Viroplasms Contain Polymerase-Associated Proteins

Martins

Johnson

Lawrence

et al. 1998

J Virol

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“…SYNV gene expression occurs through transcription of the genomic RNA into discrete mRNAs that are polyadenylated and are selectively partitioned to free and membrane-bound polysomes in the cytoplasm (35,36). In infected cells, the L mRNA is present at a lower abundance than the 3Ј-encoded mRNAs, suggesting that relative mRNA abundance could be an important component of the regulation of gene expression (29,47).…”

mentioning

confidence: 99%

Characterization of the components and activity of Sonchus yellow net rhabdovirus polymerase

Wagner

Jackson

1997

J Virol

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Sonchus yellow net virus (SYNV) is the best-characterized member of a group of plant rhabdoviruses that replicate in the host cell nucleus. Using a recently developed method for partial purification of active SYNV polymerase by salt extraction of nuclei from infected plant tissue (J. D. O. Wagner et al, J. Virol. 70:468-477, 1996), we have identified the nucleocapsid (N), M2, and L proteins as polymerase complex components (based on copurification with the polymerase activity and by coimmunoprecipitation assays). Furthermore, the L protein was shown by antibody inhibition analysis to be a functional component of the polymerase. A second complex of M2 and L proteins, thought to be a precursor to the polymerase complex, was also identified. In addition, we conducted a detailed characterization of SYNV RNA synthesis in vitro. The results demonstrate that the RNAs are transcribed sequentially, beginning with the N mRNA and followed successively by the remaining five mRNAs in the order of their genome organization. Gene expression conforms to a cascade pattern, with synthesis of the 3-proximal N mRNA occurring at the highest level, followed by consecutively lower levels of transcription from each subsequent gene. The reaction conditions favor transcription over minus-sense RNA replication, which, we posit, is inhibited near specific signal sequences located on the antigenomic template. The results support the concept that the mechanism of transcription is highly conserved among diverse rhabdoviruses and are compatible with a unified model for the regulation of genomic and antigenomic RNA synthesis.

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Morphology and development of rhabdovirus-like particles inCuscuta odorata (Convolvulaceae) simultaneous infected with virus and mycoplasma-like organisms

et al. 1987

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Subcellular distribution of RNA sequences complementary to sonchus yellow net virus RNA

Cited by 9 publications

References 12 publications

Sonchus Yellow Net Rhabdovirus Nuclear Viroplasms Contain Polymerase-Associated Proteins

Sonchus Yellow Net Rhabdovirus Nuclear Viroplasms Contain Polymerase-Associated Proteins

Characterization of the components and activity of Sonchus yellow net rhabdovirus polymerase

Morphology and development of rhabdovirus-like particles inCuscuta odorata (Convolvulaceae) simultaneous infected with virus and mycoplasma-like organisms

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