UNC-62 binds to and activates expression of all six <i>C. elegans</i> yolk protein genes.

<p>(A) The thick and thin blue lines indicate the exon and intron structure of each of the six vitellogenin loci. Red boxes indicate the position of regions significantly enriched in UNC-62 ChIP-seq of day 4 adult (YA) worms. (*) indicates <i>q</i>-value<0.001 and (**) indicates <i>q<</i>10⁻⁵ binding sites. Below each gene, RNA-seq results are displayed for worms fed either <i>unc-62</i> RNAi or control bacteria. Bars indicate the mean, and error bars the standard deviation, of sequencing read density (reads per million mapped reads) for vitellogenin genes in triplicate RNA-seq experiments. (B) Bars indicate mean, and error bars indicate standard deviation, of measured fluorescence of a <i>vit-2:GFP</i> reporter under various RNAi conditions. Expression in ∼15 worms was quantified using ImageJ. (C) (top) A <i>de novo</i> motif search in UNC-62 young adult binding sites with RSAT <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003325#pgen.1003325-ThomasChollier1" target="_blank">[58]</a> identifies a TGATTGACA motif as the prominent sequence motif. (bottom) This motif is similar to the ATTGACA VPE-1 vitellogenin regulatory motif previously described <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003325#pgen.1003325-MacMorris1" target="_blank">[30]</a>. (D) All six vitellogenin genes decrease expression with age (as assayed by whole-worm microarrays of 2, 5, 8, and 11 day old adult worms <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003325#pgen.1003325-Budovskaya1" target="_blank">[10]</a>). Bars indicate mean expression observed across replicated arrays, with error bars indicating standard deviation. (E) UNC-62 activates vitellogenin expression in old adults. Bars indicate expression of two biological replicates of <i>vit-2</i> (black) and <i>vit-6</i> (grey), determined by qRT-PCR of RNA isolated from ∼100 dissected intestines of day 1 adults and day 15 adults fed either control or <i>unc-62</i> RNAi bacteria. Error bars indicate standard deviation of triplicate qPCR technical replicates. Statistical significance is not indicated as this experiment included only two biological replicates.