Antiviral RISC mainly targets viral mRNA but not genomic RNA of tospovirus
Fig 5
RISC fraction from TZSV-infected plant targets naked genomic RNA but not the genomic RNA complexed with N proteins in vitro.
(A) and (C) In intro cleavage assay of naked genomic RNA of TZSV S and M segment by isolated RISC chromatography fractions. Chromatography fraction 5 (S6 Fig) from TZSV-infected N. benthamiana were incubated with DIG-labeled in vitro full-length RNA transcripts of TZSV S (100 ng; A) and M (100 ng; B) segment, respectively. (B) and (D) In vitro cleavage assay of TZSV genomic S or M RNA complexed with N protein by the RISC fraction. DIG-labelled full-length S (B) or M (D) genomic RNA was incubated with TZSV N protein to form N-RNA complexes. RISC fraction 5 isolated from TZSV-infected plants was added to N-S RNA or N-M RNA complexes. The blots were developed by AP-labeled anti-digoxigenin antibodies and followed with BCIP/NBT staining.