An in vitro reprogrammable antiviral RISC with size-preferential ribonuclease activity. Academic Article

abstract

• Infection of Nicotiana benthamiana plants with Tomato bushy stunt virus (TBSV) mutants compromised for silencing suppression induces formation of an antiviral RISC (vRISC) that can be isolated using chromatography procedures. The isolated vRISC sequence-specifically degrades TBSV RNA in vitro, its activity can be down-regulated by removing siRNAs, and re-stimulated by exogenous supply of siRNAs. vRISC is most effective at hydrolyzing the ~4.8kb genomic RNA, but less so for a ~2.2kb TBSV subgenomic mRNA (sgRNA1), while the 3' co-terminal sgRNA2 of ~0.9kb appears insensitive to vRISC cleavage. Moreover, experiments with in vitro generated 5' co-terminal viral transcripts show that RNAs of ~2.7kb are efficiently cleaved while those of ~1.1kb or shorter are unaffected. The isolated antiviral ribonuclease complex fails to degrade ~0.4kb defective interfering RNAs (DIs) in vitro, agreeing with findings that in plants DIs are not targeted by silencing.

authors

• Scholthof, Herman

published proceedings

• Virology

altmetric score

• 5.024

author list (cited authors)

• Omarov, R. T., Ciomperlik, J., & Scholthof, H. B.

citation count

• 6

complete list of authors

• Omarov, Rustem T||Ciomperlik, Jessica||Scholthof, Herman B

publication date

• March 2016

publisher

• Elsevier  Publisher

published in

• Virology  Journal

keywords

• Plant
• Plant Diseases
• Plant Proteins
• RNA Interference
• RNA, Viral
• Risc
• Rna Silencing
• Rna-induced Silencing Complex
• Tobacco
• Tombusvirus
• Virus

PubMed Central ID

• 26812224

Digital Object Identifier (DOI)

• 10.1016/j.virol.2015.12.017

start page

• 41

end page

• 48

volume

• 490

URL

• http%3A%2F%2Fdx.doi.org%2F10.1016%2Fj.virol.2015.12.017