Plant Viral Rna Synthesis in Cell-Free Systems

Graaff, M. de; Jaspars, E.M.J.

doi:10.1146/annurev.py.32.090194.001523

Cited by 31 publications

(18 citation statements)

References 40 publications

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“…Vesicular bodies of various sizes and shapes are cytopathological structures associated with infections of many different types of plant viruses, and in some cases these have been shown to contain viral dsRNAs and\or RNA-dependent RNA polymerases (Burgyan et al, 1996 ;Martelli et al, 1984 ;De Graaff & Jaspars, 1994 ;Lesemann, 1988). These observations and the fact that LIYV RNA 1 is alone competent for replication and primarily encodes proteins predicted to be involved in replication (Klaassen et al, 1995), further support the idea that the vesicles induced by LIYV are associated with LIYV RNA replication.…”

mentioning

confidence: 77%

Specific inclusion bodies are associated with replication of lettuce infectious yellows virus RNAs in Nicotiana benthamiana protoplasts.

Medina

Tian

Wierzchos

et al. 1998

Journal of General Virology

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Nicotiana benthamiana mesophyll protoplasts, either mock-inoculated or inoculated using in vitro transcripts derived from lettuce infectious yellows virus (LIYV) RNA 1-and/or RNA 2-cloned cDNAs were analysed by transmission electron microscopy (TEM) and, in some cases, also by immunogold labelling. TEM revealed the main cytopathological effects of LIYV infections in N. benthamiana protoplasts infected with RNAs 1 and 2 : (a) typical closterovirus-induced (beet yellows virus-type) accumulations of vesiculated cytoplasmic membranes as inclusion bodies, sometimes with associated virions ; (b) scattered aggregations of virions within the cytoplasm ; and (c) electron-dense plasmalemma deposits. These were not seen in mock-inoculated protoplasts. Protoplasts inoculated only with LIYV RNA 1 contained vesiculated cytoplasmic inclusion bodies, but not virions or plasmalemma deposits. Thus, infection by only LIYV RNA 1 is sufficient to induce characteristic closterovirus vesiculated cytoplasmic inclusion bodies. However, both LIYV RNAs 1 and 2 are needed for production of virions and plasmalemma deposits.Lettuce infectious yellows virus (LIYV) is the type member of the genus Crinivirus of the family Closteroviridae. The LIYV genome is bipartite, divided among two ssRNAs of 8118 and 7193 nucleotides (RNAs 1 and 2 respectively), giving a total of 15 311 nucleotides (Klaassen et al., 1995). On LIYV RNA 2 is the ' hallmark closterovirus gene array ' including five genes encoding : a small hydrophobic protein of as yet unknown function (ORF 1) ; an HSP70 homologue (ORF 2) ; a protein of unknown function (p59, ORF 3) ; the capsid protein (CP,

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mentioning

confidence: 77%

Specific inclusion bodies are associated with replication of lettuce infectious yellows virus RNAs in Nicotiana benthamiana protoplasts.

Medina

Tian

Wierzchos

et al. 1998

Journal of General Virology

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“…Double-stranded RNAs (usually called replicative form or RF dsRNAs) are usually found in nucleic preparations from cells infected with positive-strand RNA viruses or in the extracted products of in vitro RNA synthesis reactions with crude or partially purified polymerase preparations from infected plants or animal cells (de Graaf and Jaspars, 1994), irrespective of the polymerase or helicase type. Double-stranded RNAs (usually called replicative form or RF dsRNAs) are usually found in nucleic preparations from cells infected with positive-strand RNA viruses or in the extracted products of in vitro RNA synthesis reactions with crude or partially purified polymerase preparations from infected plants or animal cells (de Graaf and Jaspars, 1994), irrespective of the polymerase or helicase type.…”

Section: Models For Rna Replication: the Role Of Helicasesmentioning

confidence: 99%

“…In the case of flockhouse virus, it was possible to remove the bound RNA from the membrane-bound complex by nuclease digestion to produce a template-dependent RNA polymerase. For some other viruses it has been possible to produce templatedependent RNA polymerases after detergent solubilization and removal of the endogenous RNA (reviewed by de Graaf and Jaspars, 1994). It was suggested that initiation of positive-strand synthesis may result from a direct GPL-replicase interaction, analogous to activation of the E. coli replication initiator protein dnaA by diphosphatidylglycerol (Sekimizu and Kornberg, 1988), or a change in membrane configuration to mimic similar changes that may occur during replication in uiuo.…”

Section: The Role Of Membranes In Rna Replicationmentioning

confidence: 99%

Comparison of The Replication of Positive-Stranded Rna Viruses of Plants and Animals

Buck

1996

Advances in Virus Research

438

340

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“…Most purified viral RNA replication complexes copurify with membrane extracts from infected cells (reviewed in reference 10) and, although in some cases RNA synthesis activity can be solubilized (24,67), in vivo and in vitro studies suggest that the presence of membranes and/or phospholipids is essential for at least some steps of RNA replication (37,41,67). It was proposed that these membranes can play both a structural and a functional role in the replication complex.…”

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confidence: 99%

Targeting of the Turnip Yellow Mosaic Virus 66K Replication Protein to the Chloroplast Envelope Is Mediated by the 140K Protein

Prod'homme

Jakubiec

Tournier

et al. 2003

J Virol

112

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. During viral infection, the 66K protein localizes to virus-induced chloroplastic membrane vesicles, which are closely associated with TYMV RNA replication. To investigate the determinants of its subcellular localization, the 66K protein was expressed in plant protoplasts from separate plasmids. Green fluorescent protein (GFP) fusion and immunofluorescence experiments demonstrated that the 66K protein displayed a cytoplasmic distribution when expressed individually but that it was relocated to the chloroplast periphery under conditions in which viral replication occurred. The 66K protein produced from an expression vector was functional in viral replication since it could transcomplement a defective replication template. Targeting of the 66K protein to the chloroplast envelope in the course of the viral infection appeared to be solely dependent on the expression of the 140K protein. Analysis of the subcellular localization of the 140K protein fused to GFP demonstrated that it is targeted to the chloroplast envelope in the absence of other viral factors and that it induces the clumping of the chloroplasts, one of the typical cytological effects of TYMV infection. These results suggests that the 140K protein is a key organizer of the assembly of the TYMV replication complexes and a major determinant for their chloroplastic localization and retention.A universal feature of eukaryotic positive-strand RNA viruses is that replication of their genomes is closely associated with intracellular membranes (reviewed in reference 8). Most purified viral RNA replication complexes copurify with membrane extracts from infected cells (reviewed in reference 10) and, although in some cases RNA synthesis activity can be solubilized (24, 67), in vivo and in vitro studies suggest that the presence of membranes and/or phospholipids is essential for at least some steps of RNA replication (37,41,67). It was proposed that these membranes can play both a structural and a functional role in the replication complex.Electron microscopy observations of infected cells revealed that many positive-stranded RNA viruses induce proliferation and/or reorganization of the intracellular membranes of their host to create a membrane compartment in which RNA replication takes place. Depending on the virus, a variety of membrane systems can be concerned, including the early and late endomembrane systems (52, 59), the nuclear envelope (13), the vacuole (64), the endosomes and lysosomes (17, 59), the peroxisomes (56), chloroplasts (35), and mitochondria (14, 40). The fact that distinct types of membranes are involved in the replication of different viruses suggests the establishment of specific interactions between such host membranes and virusencoded proteins. A number of viral proteins that target replication complexes to intracellular membranes have been identified (9,48,55,63). Membrane interaction of host-encoded factors that are part of the viral replication complex has also been reported (22,68).Despite this universal association of positive-strand ...

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Plant Viral Rna Synthesis in Cell-Free Systems

Cited by 31 publications

References 40 publications

Specific inclusion bodies are associated with replication of lettuce infectious yellows virus RNAs in Nicotiana benthamiana protoplasts.

Specific inclusion bodies are associated with replication of lettuce infectious yellows virus RNAs in Nicotiana benthamiana protoplasts.

Comparison of The Replication of Positive-Stranded Rna Viruses of Plants and Animals

Targeting of the Turnip Yellow Mosaic Virus 66K Replication Protein to the Chloroplast Envelope Is Mediated by the 140K Protein

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