Schematics of the <i>ceh-22</i>, <i>psa-3</i> and <i>end-1</i> loci.

Abstract

<p>For each locus, black boxes represent exons and gray boxes untranslated regions (UTRs). Start codons representing the Translation Start Site (TlSS) for each isoform are marked by ‘M’. White boxes represent the genomic region used to construct the WRE reporters and the green box the GFP variant used. The larger white boxes in the WRE reporter show the location of the HMG (red lines) and Helper sites (blue lines). Below each schematic are the genomic sequences highlighting the putative Helper sites (blue) and functional HMG sites (red) that were targeted for mutagenesis. (A) For the <i>ceh-22</i> gene (Gene ID: 179485), a transcriptional fusion of the <i>ceh-22b</i> isoform called <i>ceh-22b::VENUS </i><a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1004133#pgen.1004133-Lam1" target="_blank">[42]</a> was used for reporter analysis (nucleotides −1853 to −633 with the first nucleotide of the <i>ceh-22b</i> TlSS representing +1). (B) For <i>psa-3</i> (Gene ID: 181631), a translational fusion (<i>psa-3::GFP</i>) including promoter sequences (starting at -382) and the first exons of the a, b & c isoforms was used, where the <i>pqn-36</i> gene, located in the third intron was deleted, as indicated by the parentheses <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1004133#pgen.1004133-Arata1" target="_blank">[43]</a>. (C) For <i>end-1</i> (Gene ID: 179893), a translational fusion containing ∼2.2 kb of promoter sequence, known as <i>end-1::GFP::H2B</i> was used <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1004133#pgen.1004133-Shetty1" target="_blank">[16]</a>.</p