Phosphorylation of cucumber mosaic virus RNA polymerase 2a protein inhibits formation of replicase complex

Kim, Sang Hyon; Palukaitis, Peter; Park, Young In

doi:10.1093/emboj/21.9.2292

Cited by 69 publications

(83 citation statements)

References 55 publications

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“…Othello and not elicit systemic necrosis in the RT4-4 N. benthamiana plants likely reflects the inability of RT4-4 to recognize the 2a protein of this CMV strain. Finally, a role for posttranslational modification in 2a protein recognition, by RT4-4, came from the finding that the CMV 2a phosphorylation mimic mutant failed to elicit the necrosis phenotype (37). Together, these findings are consistent with results of previous studies that identified the 2a protein as the elicitor of a CMV defense response in cowpea (22).…”

Section: Discussionsupporting

confidence: 88%

A viral resistance gene from common bean functions across plant families and is up-regulated in a non-virus-specific manner

Seo

Rojas

Lee

et al. 2006

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

103

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Genes involved in a viral resistance response in common bean (Phaseolus vulgaris cv. Othello) were identified by inoculating a geminivirus reporter (Bean dwarf mosaic virus expressing the green fluorescent protein), extracting RNA from tissue undergoing the defense response, and amplifying sequences with degenerate R gene primers. One such gene (a TIR-NBS-LRR gene, RT4-4) was selected for functional analysis in which transgenic Nicotiana benthamiana were generated and screened for resistance to a range of viruses. This analysis revealed that RT4-4 did not confer resistance to the reporter geminivirus; however, it did activate a resistance-related response (systemic necrosis) to seven strains of Cucumber mosaic virus (CMV) from pepper or tomato, but not to a CMV strain from common bean. Of these eight CMV strains, only the strain from common bean systemically infected common bean cv. Othello. Additional evidence that RT4-4 is a CMV R gene came from the detection of resistance response markers in CMV-challenged leaves of RT4-4 transgenic plants, and the identification of the CMV 2a gene product as the elicitor of the necrosis response. These findings indicate that RT4-4 functions across two plant families and is up-regulated in a non-virus-specific manner. This experimental approach holds promise for providing insights into the mechanisms by which plants activate resistance responses against pathogens.avirulence gene ͉ Cucumber mosaic virus ͉ geminivirus ͉ host defense response ͉ systemic necrosis

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Section: Discussionsupporting

confidence: 88%

A viral resistance gene from common bean functions across plant families and is up-regulated in a non-virus-specific manner

Seo

Rojas

Lee

et al. 2006

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

103

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“…This interaction has also been reported for Cucumber mosaic virus (Kim et al, 2002) and Alfalfa mosaic virus (Maurice et al, 2001), the genomes of which consist of three divided RNAs corresponding to RNAs 1, 2 and 3 of the bromoviruses, indicating that the 1a-2a interaction is a common characteristic in the family Bromoviridae. Detectable accumulation of viral RNA was directed by B1S2 and C1S2 combinations.…”

Section: Discussionsupporting

confidence: 71%

Use of Spring beauty latent virus to identify compatible interactions between bromovirus components required for virus infection

Fujisaki

Kaido

Mise

et al. 2003

Journal of General Virology

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Spring beauty latent virus (SBLV) is a member of the genus Bromovirus, and is closely related to Brome mosaic virus (BMV) and Cowpea chlorotic mottle virus (CCMV). Compatible interactions between viral components are required for successful infection of plants by BMV and CCMV. To further our understanding of interactions between bromovirus components, we used SBLV to produce reassortants among the three bromoviruses. We found that SBLV RNA 2 functioned with heterologous bromovirus RNA 1 in infections of whole plants and protoplasts of Nicotiana benthamiana, although SBLV RNA 1 did not function with heterologous bromovirus RNA 2. A DNA-based transient assay for 1a and 2a proteins, which are encoded by RNAs 1 and 2, respectively further suggested that SBLV 2a protein may function in combination with heterologous bromovirus 1a protein. Moreover, analysis of the ability of reassortants to spread locally revealed that an RNA 2-mediated interaction between viral components may be required for efficient cell-tocell movement of bromoviruses.

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“…These phosphatases might contribute to BMV replication by jointly counteracting N-terminal phosphorylation of 2a polymerase, which, for a closely related bromovirus, has been shown to occur in vivo and to inhibit 2a interaction with 1a to promote replication complex assembly (41). Thus, replication complex assembly may be regulated by cell phosphatases as well as kinases.…”

Section: Discussionmentioning

confidence: 99%

Systematic, genome-wide identification of host genes affecting replication of a positive-strand RNA virus

Kushner

Lindenbach²,

Grdzelishvili³

et al. 2003

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

236

250

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Brome mosaic virus (BMV), a member of the alphavirus-like superfamily of human-, animal-, and plant-infecting (ϩ)RNA viruses, has been studied as a model for viral RNA replication, encapsidation, recombination, and other processes (3). BMV has three genomic RNAs. RNAs 1 and 2 encode the interacting, multifunctional 1a helicase-like and 2a polymerase RNA replication factors (4, 5), which form endoplasmic reticulum (ER) membrane-associated RNA replication complexes with functional similarities to the replicative cores of retrovirus and double-strand (ds)RNA virus virions (6). RNA3 encodes protein 3a that enables infection spread between cells in natural hosts. The negative-strand [(Ϫ) RNA]3 replication intermediate also serves as a template for synthesis of a subgenomic (sg) mRNA, RNA4, which encodes the viral coat protein (Fig. 1A).The yeast Saccharomyces cerevisiae has proven a valuable model for normal and disease processes in human and other cells. The unusual ability of BMV to direct its genomic RNA replication, gene expression, encapsidation, and other processes in this yeast (7,8) has allowed traditional yeast mutagenic analyses that have identified host genes involved in multiple steps of BMV RNA replication and gene expression. Such host genes encode a wide variety of functions and contribute to diverse replication steps, including supporting and regulating viral translation, selecting and recruiting viral RNAs as replication templates, activating the RNA replication complex through chaperones, and providing a lipid profile compatible with membrane-associated viral RNA replication (9-14; reviewed in refs. 2 and 15).Here, we sought to develop a more rapid, global method to systematically identify yeast host factors with effects on BMV RNA replication by using an ordered array of yeast deletion strains (16) to assay virus replication in the absence of each of Ϸ4,500 yeast factors, which is Ϸ80% of the yeast genome. We describe screening this deletion array by using a whole-cell assay based on BMV-directed Renilla luciferase (Rluc) expression by pathways dependent on viral RNA replication and viral RNAdirected sg mRNA synthesis. The assay identified nearly 100 host genes whose absence repressed or enhanced BMV-directed Rluc expression by 3-to 25-fold. The results provide a significantly expanded view of virus-host interactions and should advance understanding of virus and cell pathways. Materials and MethodsYeast. YMI04 and ded1i yeast were described (11). Strains BY4743 (WT; ref. 17) and the homozygous diploid deletion series (BY4743 strain background; ref. 16) were from Research Genetics (Huntsville, AL). Standard yeast techniques were used (18), except for 96-well transformations, which were based on a one-step procedure (19). Briefly, yeast were grown to saturation overnight at 30°C in 96-well plates (1.2 ml per well), pelleted, suspended in 100 l of transformation mix (0.18 M LiAc, pH 5.5, 36% polyethylene glycol-3350, 90 mM DTT, 0.5 mg͞ml sheared salmon sperm DNA, and 20 g͞ml of each plasmid), incubate...

show abstract

Phosphorylation of cucumber mosaic virus RNA polymerase 2a protein inhibits formation of replicase complex

Cited by 69 publications

References 55 publications

A viral resistance gene from common bean functions across plant families and is up-regulated in a non-virus-specific manner

A viral resistance gene from common bean functions across plant families and is up-regulated in a non-virus-specific manner

Use of Spring beauty latent virus to identify compatible interactions between bromovirus components required for virus infection

Systematic, genome-wide identification of host genes affecting replication of a positive-strand RNA virus

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