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oai:figshare.com:article/34049533 2026-10-01T17:47:03Z category_4 category_12 category_13 category_14 category_15 category_16 category_64 category_128 portal_5 item_type_1 month_year_10_2026

egl-2(G574E) is a gain-of-function allele. Xinyu Huang (679541) Sudharsan Kannan (25158064) Gail A. Robertson (15007385) Han Wang (254423) Biochemistry Cell Biology Genetics Molecular Biology Neuroscience Physiology Cancer Hematology regulate neuronal excitability promoting neuron ala influence channel gating hallmark cyclic nucleotide genetic analysis demonstrated forward genetics reveals electrophysiological analysis showed dependent channel activation calcium imaging showed binding homology domains adjacent channel subunits accelerates activation kinetics subfamily kcnh channels conserved gating mechanism important sleep regulator findings identify egl div >< p caenorhabditis elegans </ similarly suppresses sleep 2 functions cell elegans </ suppresses sleep kcnh voltage vivo </ conserved cnbhd central sleep shifts voltage second substitution negative potentials intrinsic ligand g574r ), function effects corresponding residue (A) Gene structure of egl-2 showing the positions and the mutations of the indicated alleles. gof, gain-of-function; lof, loss-of-function. Open shapes, gray rectangles, and solid lines indicate UTRs, exons, and introns of egl-2 isoform a, respectively. egl-2(n2656[A478V]) is a well-characterized gain-of-function allele. egl-2(gk593448) carries a G574R mutation at the same glycine residue mutated in egl-2(tan200[G574E]). egl-2(tan232) is a loss-of-function allele with the entire egl-2 gene deleted. STOP-IN is a 43-bp cassette that contains multiple stop codons in all three reading frames, which disrupts gene function when it is inserted into coding sequences [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1012255#pgen.1012255.ref031" target="_blank">31</a>]. tan209, tan325, and tan323 were generated in the egl-2(tan200) mutant background using CRISPR. Some of these alleles were used in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1012255#pgen.1012255.g005" target="_blank">Figs 5</a> and <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1012255#pgen.1012255.g006" target="_blank">6</a>. (B) Time course of endogenous stress-induced sleep in animals of the indicated genotypes, plotted as the fraction of quiescent worms at each time point. Pre refers to before heat shock. Data were from three independent replicates, with 15–25 animals of each strain assayed on separate plates in each replicate. Mean ± SEM are shown. (C-E) Quantification of sleep behavior shown as the fraction of quiescent worms for the indicated genotypes and conditions. All strains contain the transgene (HS::EGF) for conditional overexpression of EGF via 15-min heat shock (HS) at 33 °C to induce sleep. Pre and post refer to before and 2 h after heat shock, respectively. (C) The phenotype of egl-2(tan200) is similar to that of egl-2(n2656, gof). (D) egl-2(tan200tan209[STOP-IN]) carries a STOP-IN cassette that disrupts the function of endogenous egl-2(tan200[G574E]). (E) egl-2(G574E) or egl-2(+) cDNA sequences were overexpressed in the ALA neuron of otherwise wild-type animals using an ALA-specific promoter (Pnlp-67). Each dot represents one independent assay containing 15–25 animals on a single plate. Mean ± SEM are shown. ns, not significant; *, p < 0.05; ****, p < 0.0001. One-way ANOVA with Dunnett’s multiple-comparisons test for post-heat shock samples, comparing each genotype with egl-2(tan200) (C-D) or wild type (E). 2026-10-01T17:54:06Z Image Figure 10.1371/journal.pgen.1012255.g004 https://figshare.com/articles/figure/_p_i_egl-2_G574E_i_is_a_gain-of-function_allele_p_/34049533 CC BY 4.0