RNA-dependent RNA polymerases in uninfected and tobacco mosaic virus-infected tobacco leaves: Viral-induced stimulation of a host polymerase activity

Romaine, C. P.; Zaitlin, Milton

doi:10.1016/0042-6822(78)90024-7

Cited by 68 publications

(20 citation statements)

References 25 publications

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“…In several studies, elevated RdRP activity was found in virus-infected plants and has been speculated to be one of the host factors involved in virus replication (5,(8)(9)(10). In the present study, we have shown that RdRP activity is induced not only by virus infection but also by defense-inducing compounds (Fig.…”

Section: Discussionsupporting

confidence: 61%

“…In these assays, we detected synthesis of not only the high molecular weight TMV RNAs by TMV replicase but also smaller RNAs by an unknown RNA polymerase. Synthesis of smaller RNAs has been previously observed in similar assays and attributed to the activity of a plant RdRP (5,(8)(9)(10). Interestingly, in our assays, synthesis of these smaller RNAs was drastically increased in SA-treated tobacco plants (data not shown).…”

Section: Sa Induces a Rdrp Activity In Tobaccosupporting

confidence: 55%

“…R NA-dependent RNA polymerase (RdRP) activities have been detected in a number of plants (1)(2)(3)(4)(5)(6)(7)(8)(9)(10). Recently, a tomato RdRP has been purified and its corresponding gene cloned (11)(12)(13).…”

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confidence: 99%

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An important role of an inducible RNA-dependent RNA polymerase in plant antiviral defense

Xie

Fan²,

Chen³

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

282

254

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Plants contain RNA-dependent RNA polymerase (RdRP) activities that synthesize short cRNAs by using cellular or viral RNAs as templates. During studies of salicylic acid (SA)-induced resistance to viral pathogens, we recently found that the activity of a tobacco RdRP was increased in virus-infected or SA-treated plants. Biologically active SA analogs capable of activating plant defense response also induced the RdRP activity, whereas biologically inactive analogs did not. A tobacco RdRP gene, NtRDRP1, was isolated and found to be induced both by virus infection and by treatment with SA or its biologically active analogs. Tobacco lines deficient in the inducible RDRP activity were obtained by expressing antisense RNA for the NtRDRP1 gene in transgenic plants. When infected by tobacco mosaic virus, these transgenic plants accumulated significantly higher levels of viral RNA and developed more severe disease symptoms than wild-type plants. After infection by a strain of potato virus X that does not spread in wild-type tobacco plants, the transgenic NtRDRP1 antisense plants accumulated virus and developed symptoms not only locally in inoculated leaves but also systemically in upper uninoculated leaves. These results strongly suggest that inducible RdRP activity plays an important role in plant antiviral defense.

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Section: Discussionsupporting

confidence: 61%

Section: Sa Induces a Rdrp Activity In Tobaccosupporting

confidence: 55%

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An important role of an inducible RNA-dependent RNA polymerase in plant antiviral defense

Xie

Fan²,

Chen³

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

282

254

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“…Replicase has been extracted from smooth membranes of encephalomyocarditis virus-infected BHK-21 hamster cells (4), but BHK rRNA and Qf3 RNAs were equally good templates. RNA-dependent RNA polymerase activity has been described for plant tissues infected with several viruses, including tobacco mosaic virus (TMV) (5,6), turnip yellow mosaic virus (7), alfalfa mosaic virus (8), tobacco necrosis virus (9), cucumber mosaic virus (10), cowpea chlorotic mottle virus (CCMV) (11), and brome mosaic virus (BMV) (12)(13)(14). However, in most of these instances, stimulation of RNA polymerase activity can also be detected in mock-infected (wounded) tissue. We describe here the extraction and high biological activity of an enzyme from BMV-infected barley leaves that is considerably stimulated by the addition of RNA and for which BMV RNA is a markedly better template than other RNAs tested thus far.…”

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confidence: 99%

Highly active template-specific RNA-dependent RNA polymerase from barley leaves infected with brome mosaic virus.

Hardy¹,

German²,

Loesch-Fries³

et al. 1979

Proc. Natl. Acad. Sci. U.S.A.

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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 enzyme was more than 50 times more active than was a comparable preparation from mock-inoculated leaves and was stimulated more than 15-fold by the addition of BMV RNA to the reaction. Other viral RNA templates were less than 25% as efficient as was BMV RNA in stimulating UMP incorporation; poly(A), tRNA, and mRNA gave little stimulation and rRNA was inactive. Autoradiographic analysis after electrophoretic separation of the radioactive products from reaction mixtures containing BMV RNA template revealed prominent bands that coelectrophoresed with replicative forms of BMV RNAs. When BMV RNA template was enriched in RNA3 or RNA4, larger proportions of the products were replicative forms of RNA3 or RNA4, respectively.

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“…Brinton, 1986), which is stimulated by virus infection (Romaine & Zaitlin, 1978;Gordon et aL, 1982;Dorssers et al, 1983), usually hampers the study of viral R N A replicases involved in virus R N A replication. In fact, the level of [32p]UMP incorporation in trichloroacetic acidinsoluble material was so high in healthy N. clevelandii leaf extracts that R N A replicase activity induced by PPV infection could not be detected at this stage.…”

Section: P P V R N a Replication 789mentioning

confidence: 99%

Plum pox potyvirus RNA replication in a crude membrane fraction from infected Nicotiana clevelandii leaves

Martín

Garcı́a

1991

Journal of General Virology

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In vitro synthesis of plum pox potyvirus (PPV)-specific nucleic acid has been measured in a crude fraction prepared from leaves of PPV-infected Nicotiana clevelandii plants. Using alkali and DNase treatments, the synthesized nucleic acid was shown to be RNA. The electrophoretic mobility and the differing sensitivity to RNase at high and low salt concentrations allowed the identification of in vitro products probably corresponding to replicative form and replicative intermediate RNA, as well as to single-stranded RNA. Most of the PPV-specific RNA synthesized was shown to be of positive polarity. The in vitro RNA synthesis, performed in the presence of actinomycin D, required all four ribonucleoside triphosphates and Mg 2+ ions. This enzyme extract contained about 6 % of the leaf protein and most of the identified virus-encoded proteins.Altogether, the results presented in this paper suggest that in vitro RNA synthesis was carried out by the PPV replicase complex.

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RNA-dependent RNA polymerases in uninfected and tobacco mosaic virus-infected tobacco leaves: Viral-induced stimulation of a host polymerase activity

Cited by 68 publications

References 25 publications

An important role of an inducible RNA-dependent RNA polymerase in plant antiviral defense

An important role of an inducible RNA-dependent RNA polymerase in plant antiviral defense

Highly active template-specific RNA-dependent RNA polymerase from barley leaves infected with brome mosaic virus.

Plum pox potyvirus RNA replication in a crude membrane fraction from infected Nicotiana clevelandii leaves

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