A key antisense sRNA modulates the oxidative stress response and virulence in Xanthomonas oryzae pv. oryzicola
Fig 2
Analysis of the Xonc3711-xoc_3982 RNA interaction.
(a) Nucleotide sequence of BLS256 xoc_3982 mRNA from its initiation codon (+3) to +1245 in the CDS, with those of the predicted export signal sequence in blue font. Blue panel: The location of point mutations in Xoc_3982*; Green panel: The location of point mutations in Xonc3711*. (b) Schematic showing Xoc_3982::gfp translational fusions. Two fusions were constructed that varied in the number of nucleotides in the coding sequence. Constructs X+3 and X+1242 contained three and 1242 nucleotides of the Xoc_3982 coding sequence fused to GFP, respectively. (c) Northern blot showing Xonc3711 expression levels in the presence of reporter constructs X+3 and X+1242 (upper two panels). The lower two panels show Xoc_3982GFP proteins detected by western blotting. GFP fusion proteins were detected using antibodies directed against GFP (1:2000, mouse anti-GFP; Roche). (d) Interaction of Xonc3711 sRNA and full-length xoc_3982 mRNA in electrophoretic mobility shift assays. Lane 1, biotinylated Xonc3711; lane 2, biotinylated Xonc3711 and 40 μM xoc_3982 mRNA; lane 3, biotinylated Xonc3711 and 20 μM xoc_3982 mRNA; and lane 4, biotinylated Xonc3711, xoc_3982 mRNA, and unlabeled Xonc3711.