Nuclear localization of Beet black scorch virus capsid protein and its interaction with importin α

Zhang, Yanjing; Zhang, Xiaofeng; Niu, Shaofang; Han, Chenggui; Yu, Jialin; Li, Dawei

doi:10.1016/j.virusres.2010.10.029

Cited by 26 publications

(27 citation statements)

References 58 publications

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“…The PCR products were digested with Xba I/ Kpn I and inserted into the corresponding sites of pSuper1300-GFP. The constructed pSuper1300-LTP-GFP was used for Agrobacterium -mediated transient transformation of tobacco epidermal cells (Zhang Y. et al, 2010). pSuper1300-GFP was used as the control.…”

Section: Methodsmentioning

confidence: 99%

“…LTPs occur in the plasma membrane (Debono et al, 2009; Kim et al, 2012), cell wall (Thoma et al, 1993), or cytoplasm (Guo et al, 2013; Edstam et al, 2014). LTPs are reported to probable participated in cutin synthesis (Pyee et al, 1994; Han et al, 2001; Debono et al, 2009; Kim et al, 2012), pathogen defense responses (Maldonado et al, 2002; Silverstein et al, 2007; Guo et al, 2013; Yu et al, 2013), reproductive development (Chae et al, 2009; Zhang D. et al, 2010; Zhang Y. et al, 2010), and adaption to abiotic stresses (Guo et al, 2013; Pitzschke et al, 2014), even though their functions remain unclear.…”

Section: Introductionmentioning

confidence: 99%

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A Non-specific Setaria italica Lipid Transfer Protein Gene Plays a Critical Role under Abiotic Stress

Pan

Jiao

et al. 2016

Front. Plant Sci.

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Lipid transfer proteins (LTPs) are a class of cysteine-rich soluble proteins having small molecular weights. LTPs participate in flower and seed development, cuticular wax deposition, also play important roles in pathogen and abiotic stress responses. A non-specific LTP gene (SiLTP) was isolated from a foxtail millet (Setaria italica) suppression subtractive hybridization library enriched for differentially expressed genes after abiotic stress treatments. A semi-quantitative reverse transcriptase PCR analysis showed that SiLTP was expressed in all foxtail millet tissues. Additionally, the SiLTP promoter drove GUS expression in root tips, stems, leaves, flowers, and siliques of transgenic Arabidopsis. Quantitative real-time PCR indicated that the SiLTP expression was induced by NaCl, polyethylene glycol, and abscisic acid (ABA). SiLTP was localized in the cytoplasm of tobacco leaf epidermal cells and maize protoplasts. The ectopic expression of SiLTP in tobacco resulted in higher levels of salt and drought tolerance than in the wild type (WT). To further assess the function of SiLTP, SiLTP overexpression (OE) and RNA interference (RNAi)-based transgenic foxtail millet were obtained. SiLTP-OE lines performed better under salt and drought stresses compared with WT plants. In contrast, the RNAi lines were much more sensitive to salt and drought compared than WT. Electrophoretic mobility shift assays and yeast one-hybrids indicated that the transcription factor ABA-responsive DRE-binding protein (SiARDP) could bind to the dehydration-responsive element of SiLTP promoter in vitro and in vivo, respectively. Moreover, the SiLTP expression levels were higher in SiARDP-OE plants compared than the WT. These results confirmed that SiLTP plays important roles in improving salt and drought stress tolerance of foxtail millet, and may partly be upregulated by SiARDP. SiLTP may provide an effective genetic resource for molecular breeding in crops to enhance salt and drought tolerance levels.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A Non-specific Setaria italica Lipid Transfer Protein Gene Plays a Critical Role under Abiotic Stress

Pan

Jiao

et al. 2016

Front. Plant Sci.

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“…Comparative analysis of transcriptomic and proteomic responses of N. benthamiana –virus interactions have been performed [40], [41], [42], [43] and a microarray platform suitable for RNA profiling of virus-infected N. benthamiana (the Nb-array) is also available [2]. Although a large number of differently-expressed genes have been identified using the methods described above, more accurate quantitative analysis of target genes, in order to further confirm their transcription levels within particular contexts, is absolutely necessary before taking the next step.…”

Section: Introductionmentioning

confidence: 99%

Validation of Reference Genes for Gene Expression Studies in Virus-Infected Nicotiana benthamiana Using Quantitative Real-Time PCR

et al. 2012

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Nicotiana benthamiana is the most widely-used experimental host in plant virology. The recent release of the draft genome sequence for N. benthamiana consolidates its role as a model for plant–pathogen interactions. Quantitative real-time PCR (qPCR) is commonly employed for quantitative gene expression analysis. For valid qPCR analysis, accurate normalisation of gene expression against an appropriate internal control is required. Yet there has been little systematic investigation of reference gene stability in N. benthamiana under conditions of viral infections. In this study, the expression profiles of 16 commonly used housekeeping genes (GAPDH, 18S, EF1α, SAMD, L23, UK, PP2A, APR, UBI3, SAND, ACT, TUB, GBP, F-BOX, PPR and TIP41) were determined in N. benthamiana and those with acceptable expression levels were further selected for transcript stability analysis by qPCR of complementary DNA prepared from N. benthamiana leaf tissue infected with one of five RNA plant viruses (Tobacco necrosis virus A, Beet black scorch virus, Beet necrotic yellow vein virus, Barley stripe mosaic virus and Potato virus X). Gene stability was analysed in parallel by three commonly-used dedicated algorithms: geNorm, NormFinder and BestKeeper. Statistical analysis revealed that the PP2A, F-BOX and L23 genes were the most stable overall, and that the combination of these three genes was sufficient for accurate normalisation. In addition, the suitability of PP2A, F-BOX and L23 as reference genes was illustrated by expression-level analysis of AGO2 and RdR6 in virus-infected N. benthamiana leaves. This is the first study to systematically examine and evaluate the stability of different reference genes in N. benthamiana. Our results not only provide researchers studying these viruses a shortlist of potential housekeeping genes to use as normalisers for qPCR experiments, but should also guide the selection of appropriate reference genes for gene expression studies of N. benthamiana under other biotic and abiotic stress conditions.

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“…Analysis of TNV virion structure identified an N-terminal basic region in its CP that is disordered and oriented toward the inside of the particle [14]. Similar basic regions were also found in the CPs of other necroviruses, including TNV-D [15], Olive latent virus 1 (OLV-1) [16], TNV-A [17,18], and Beet black scorch virus (BBSV) [19]. However, the functional role of basic amino acid residues in this region is still poorly understood.…”

Section: Introductionmentioning

confidence: 99%

N-terminal basic amino acid residues of Beet black scorch virus capsid protein play a critical role in virion assembly and systemic movement

Zhang

Zhao

Zhang

et al. 2013

Virol J

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BackgroundBeet black scorch virus (BBSV) is a small single-stranded, positive-sense RNA plant virus belonging to the genus Necrovirus, family Tombusviridae. Its capsid protein (CP) contains a 13 amino acid long basic region at the N-terminus, rich in arginine and lysine residues, which is thought to interact with viral RNA to initiate virion assembly.ResultsIn the current study, a series of BBSV mutants containing amino acid substitutions as well as deletions within the N-terminal region were generated and examined for their effects on viral RNA replication, virion assembly, and long distance spread in protoplasts and whole host plants of BBSV. The RNA-binding activities of the mutated CPs were also evaluated in vitro. These experiments allowed us to identify two key basic amino acid residues in this region that are responsible for initiating virus assembly through RNA-binding. Proper assembly of BBSV particles is in turn needed for efficient viral systemic movement.ConclusionsWe have identified two basic amino acid residues near the N-terminus of the BBSV CP that bind viral RNA with high affinity to initiate virion assembly. We further provide evidence showing that systemic spread of BBSV in infected plants requires intact virions. This study represents the first in-depth investigation of the role of basic amino acid residues within the N-terminus of a necroviral CP.

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Nuclear localization of Beet black scorch virus capsid protein and its interaction with importin α

Cited by 26 publications

References 58 publications

A Non-specific Setaria italica Lipid Transfer Protein Gene Plays a Critical Role under Abiotic Stress

A Non-specific Setaria italica Lipid Transfer Protein Gene Plays a Critical Role under Abiotic Stress

Validation of Reference Genes for Gene Expression Studies in Virus-Infected Nicotiana benthamiana Using Quantitative Real-Time PCR

N-terminal basic amino acid residues of Beet black scorch virus capsid protein play a critical role in virion assembly and systemic movement

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