Structure of Hibiscus Latent Singapore Virus by Fiber Diffraction: A Nonconserved His122 Contributes to Coat Protein Stability

Tewary, Sunil Kumar; Oda, Tatsuya; Kendall, Amy; Bian, Wen; Stubbs, Gerald; Wong, Sek‐Man; Swaminathan, Kunchithapadam

doi:10.1016/j.jmb.2010.12.032

Cited by 8 publications

(7 citation statements)

References 47 publications

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“…When the seedlings reached 6 to 8 fully expanded leaves, they were inoculated with mock inoculation buffer (0.01M phosphate buffer, pH 7.0) or HLSV by mechanical inoculation. Two leaves on each plant were dusted with carborundum and inoculated with mock buffer (negative control) or purified HLSV [62]. Similarly, two leaves on each plant were inoculated with TMV.…”

Section: Methodsmentioning

confidence: 99%

Profiling of Genes Related to Cross Protection and Competition for NbTOM1 by HLSV and TMV

2013

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Cross protection is the phenomenon through which a mild strain virus suppresses symptoms induced by a closely related severe strain virus in infected plants. Hibiscus latent Singapore virus (HLSV) and Tobacco mosaic virus (TMV) are species within the genus tobamovirus. HLSV can protect Nicotiana benthamiana against TMV-U1 strain, resulting in mild symptoms instead of severe systemic necrosis. The mechanism of cross protection between HLSV and TMV is unknown. In the past, some researchers suggest that the protecting virus strain might occupy virus-specific replication sites within a cell leaving no room for the challenge virus. Quantitative real-time RT-PCR was performed to detect viral RNA levels during cross protection. HLSV accumulation increased in cross protected plants compared with that of single HLSV infected plants, while TMV decreased in cross protected plants. This suggests that there is a competition for host factors between HLSV and TMV for replication. To investigate the mechanism under the cross protection between HLSV and TMV, microarray analysis was conducted to examine the transcriptional levels of global host genes during cross protection, using Tobacco Gene Expression Microarray, 4x44 k slides. The transcriptional level of some host genes corresponded to accumulation level of TMV. Some host genes were up-regulated only by HLSV. Tobamovirus multiplication gene 1 (TOM1), essential for tobamovirus multiplication, was involved in competition for replication by HLSV and TMV during cross protection. Both HLSV and TMV accumulation decreased when NbTOM1 was silenced. A large quantity of HLSV resulted in decreased TMV accumulation in HLSV+TMV (100:1) co-infection. These results indicate that host genes involved in the plant defense response and virus multiplication are up-regulated by challenge virus TMV but not by protecting virus HLSV during cross protection.

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Section: Methodsmentioning

confidence: 99%

Profiling of Genes Related to Cross Protection and Competition for NbTOM1 by HLSV and TMV

2013

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“…The supernatants were examined for the presence of virions under FEI Tecnai T12 transmission electron microscope (TEM) by negative staining of samples with 2% uranyl acetate (Zechmann and Zellnig, 2009). To inoculate purified virions onto N. benthamiana, virions from TMV and TMV-43A inoculated plants were partially purified by low speed and ultra-high speed centrifugation (Tewary et al, 2011).…”

Section: Hlsv Rna Stability Test In N Benthamiana Protoplastsmentioning

confidence: 99%

The length of an internal poly(A) tract of hibiscus latent Singapore virus is crucial for its replication

et al. 2015

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Hibiscus latent Singapore virus (HLSV) mutants were constructed to study roles of its internal poly(A) tract (IPAT) in viral replication and coat protein (CP) expression. Shortening of the IPAT resulted in reduced HLSV RNA accumulation and its minimal length required for HLSV CP expression in plants was 24 nt. Disruption of a putative long range RNA-RNA interacting structure between 5' and 3' untranslated regions of HLSV-22A and -24A resulted in reduced viral RNA and undetectable CP accumulation in inoculated leaves. Replacement of the IPAT in HLSV with an upstream pseudoknot domain (UPD) of Tobacco mosaic virus (TMV) or insertion of the UPD to the immediate downstream of a 24 nt IPAT in HLSV resulted in drastically reduced viral RNA replication. Plants infected with a TMV mutant by replacement of the UPD with 43 nt IPAT exhibited milder mosaic symptoms without necrosis. We have proposed a model for HLSV replication.

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“…Hibiscus latent Singapore virus (HLSV) is a helical rod virus approximately 300nM in length with the ssRNA genome packaged in a small channel at its center [3]. It is in the Tobamovirus family along with tobacco mosaic virus (TMV) and their symmetry and structure determination by X-ray fiber diffraction set them apart from spherical capsid structures.…”

Section: Rna Virusesmentioning

confidence: 99%

Nucleic acid packaging in viruses

Speir

Johnson

2012

Current Opinion in Structural Biology

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We review recent literature describing protein nucleic acid interactions and nucleic acid organization in viruses. The nature of the viral genome determines its overall organization and its interactions with the capsid protein. Genomes composed of single strand (ss) RNA and DNA are highly flexible and, in some cases, adapt to the symmetry of the particle-forming protein to show repeated, sequence independent, nucleoprotein interactions. Genomes composed of double stranded (ds) DNA do not interact strongly with the container due to their intrinsic stiffness, but form well-organized layers in virions. Assembly of virions with ssDNA and ssRNA genomes usually occurs through a cooperative condensation of the protein and genome, while dsDNA viruses usually pump the genome into a preformed capsid with a strong, virally encoded, molecular motor complex. We present data that suggests the packing density of ss genomes and ds genomes are comparable, but the latter exhibit far higher pressures due to their stiffness.

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Structure of Hibiscus Latent Singapore Virus by Fiber Diffraction: A Nonconserved His122 Contributes to Coat Protein Stability

Cited by 8 publications

References 47 publications

Profiling of Genes Related to Cross Protection and Competition for NbTOM1 by HLSV and TMV

Profiling of Genes Related to Cross Protection and Competition for NbTOM1 by HLSV and TMV

The length of an internal poly(A) tract of hibiscus latent Singapore virus is crucial for its replication

Nucleic acid packaging in viruses

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