Intervention of geminiviral replication in yeast by ribozyme mediated downregulation of its Rep protein

Chilakamarthi, Ushasri; Mukherjee, Sunil Kumar; Deb, J. K.

doi:10.1016/j.febslet.2007.04.084

Cited by 11 publications

(16 citation statements)

References 31 publications

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“…The catalytic efficiency, i.e. the turnover number k cat /K M , was 0.31 10 6 min -1 M -1 , a value comparable with those described for other biologically active ribozymes [18,19]. …”

Section: Resultssupporting

confidence: 79%

Ribozyme-mediated gene knock down strategy to dissect the consequences of PDGF stimulation in vascular smooth muscle cells

et al. 2012

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BackgroundVascular Smooth Muscle Cells (VSMCs), due to their plasticity and ability to shift from a physiological contractile-quiescent phenotype to a pathological proliferating-activated status, play a central role in the onset and progression of atherosclerosis and cardiovascular diseases. PDGF-BB, among a series of cytokines and growth factors, has been identified as the critical factor in this phenotypic switch. In order to obtain new insights on the molecular effects triggered by PDGF-BB, a hammerhead ribozyme targeting the membrane receptor PDGFR-β was applied to inhibit PDGF pathway in porcine VSMCs.FindingsRibozymes, loaded on a cationic polymer-based vehicle, were delivered into cultured VSMCs. A significant impairment of the activation mechanisms triggered by PDGF-BB was demonstrated since cell migration decreased after treatments. In order to functionally validate the effects of PDGFR-β partial knock down we focused on the phosphorylation status of two proteins, protein disulfide isomerase-A3 (PDI-A3) and heat shock protein-60 (HSP-60), previously identified as indicative of VSMC phenotypic switch after PDGF-BB stimulation. Interestingly, while PDI-A3 phosphorylation was counteracted by the ribozyme administration indicating that PDI-A3 is a factor downstream the receptor signalling cascade, the HSP-60 phosphorylation status was greatly increased by the ribozyme administration.ConclusionThese contradictory observations suggested that PDGF-BB might trigger different parallel pathways that could be modulated by alternative isoforms of the receptors for the growth factor. In conclusion the knock down strategy here described enables to discriminate between two tightly intermingled pathways. Moreover it opens new attractive perspectives in functional investigations where combined gene knock down and proteomic technologies would allow the identification of key factors and pathways involved in VSMC-linked pathological disorders.

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“…The catalytic efficiency, i.e. the turnover number k cat /K M , was 0.31 10 6 min -1 M -1 , a value comparable with those described for other biologically active ribozymes [18,19]. …”

Section: Resultssupporting

confidence: 79%

Ribozyme-mediated gene knock down strategy to dissect the consequences of PDGF stimulation in vascular smooth muscle cells

et al. 2012

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“…In the first report on this strategy, it was revealed that the hammerhead ribozyme was targeted to the mRNA of replication initiator protein (Rep) of Mungbean yellow mosaic India virus (MYMIV), ultimately causing a significant decrease in MYMIV replication (Fig. 1) [7]. Following this, targeting other necessary geminiviral proteins can provide possibilities for superior virus resistance in plants.…”

Section: Role Of Ribozyme Mediated Strategy In Hindering Viral Replicmentioning

confidence: 98%

Application of molecular antiviral compounds: novel approach for durable resistance against geminiviruses

Sahu

Prasad

2015

Mol Biol Rep

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Both transgenic as well as traditional breeding approaches have not been completely successful in inducting resistance against geminiviruses in crop plants. This demands the utilization of non-viral, non-plant compounds possessing antiviral characteristics as an alternate and effective strategy for developing durable resistance against geminiviruses. In recent years, several antiviral molecules have been developed for the treatment of plant virus infections. These molecular antiviral compounds target various geminiviral-DNA and -protein via interacting with them or by cleaving viral RNA fragments. Applications of these proteins such as GroEL, g5g and VirE2 have also provided a convincing evidence of resistance against geminiviruses. Taking advantage of this information, we can generate robust resistance against geminiviruses in diverse crop plants. In this context, the present review provides epigrammatic information on these antiviral compounds and their mode of action in modulating virus infection.

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“…Rep is the most important viral protein for DNA replication. In further study with MYMIV, Chilakamarthi et al (2007) designed an anti-Rep ribozyme to target viral DNA replication. When evaluated in S. cerevisiae W303a, which lacks RNAi machinery and is thus a suitable surrogate host (Raghavan et al 2004) for replication of the virus DNA, in the presence of the anti-Rep ribozyme the yeast cell, grew slower because its growth was dependent on the begomovirus replication.…”

Section: Mungbean Yellow Mosaic Diseasementioning

confidence: 99%

Significant plant virus diseases in India and a glimpse of modern disease management technology

Rishi

2009

J Gen Plant Pathol

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Intervention of geminiviral replication in yeast by ribozyme mediated downregulation of its Rep protein

Cited by 11 publications

References 31 publications

Ribozyme-mediated gene knock down strategy to dissect the consequences of PDGF stimulation in vascular smooth muscle cells

Ribozyme-mediated gene knock down strategy to dissect the consequences of PDGF stimulation in vascular smooth muscle cells

Application of molecular antiviral compounds: novel approach for durable resistance against geminiviruses

Significant plant virus diseases in India and a glimpse of modern disease management technology

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