Figure 3: DsrA decreases the longevity of C. elegans by targeting F42G9.6.
From: Escherichia coli noncoding RNAs can affect gene expression and physiology of Caenorhabditis elegans
(a) Structure of DsrA; boxed sequence is similar to F42G9.6 mRNA (up). Identical sequence in DsrA and F42G9.6 mRNA (bottom); nucleotide of F42G9.6 mRNA is numbered from the start codon. (b) N2 had increased longevity on DsrA mutant strains than on the wild-type K12. (c) Compared with N2 worms, F42G9.6(tm2908) worms had decreased longevity on K12, and showed no significant difference on longevity when fed with K12 or DsrA mutant. tm3026 gave similar results as tm2908, and data were not shown. N=45 in b,c. (d) F42G9.6 feeding RNAi decreased longevity in both N2 and daf-2 worms, and daf-2 worms showed more significant changes to F42G9.6 feeding RNAi. N=120. (e) Overexpressing of F42G9.6 with a transgene increased C. elegans longevity. Data were from two Pdpy-30::F42G9.6::gfp lines, F42G9.6(1) and F42G9.6(2). N=95 (Pdpy-30::gfp), 86 (F42G9.6(1)) and 44 (F42G9.6(2)). (f) Semi-quantitative PCR (left) and real-time PCR (right) showed increased F42G9.6 mRNA levels in N2 worms feeding on DsrA deletion strains. (g) Representative images (up) to show that injection of the OxyS RNA decreased GFP expression in Pdpy-30::F42G9.6::gfp worms. A modified DsrA containing a scrambled sequence in the region predicted to target F42G9.6 (DsrA-NC) was used as a negative control. GFP images were taken with the same parameters. Relative quantifications of GFP signals from 20 worms were shown (bottom). Scale bar is 50 μm. (h) Semi-quantitative PCR (up) and real-time PCR (bottom) to show increased F42G9.6 mRNA levels in daf-2 worms.(i) F42G9.6 feeding RNAi decreased survival rates in both N2 and daf-2 worms when challenged with paraquat to generate oxidative stress, and daf-2 worms showed more significant changes to F42G9.6 feeding RNAi. N=120 for each group. The log-rank test was used to calculate P values for longevity and survival assays. P values were determined by two-tailed Student's t-test for the other data. *P<0.05; **P<0.01. All data are from three repeats. Error bars represent s.e.m.
