Fig. 1

The zebrafish b1115 mutation has a non-synonymous single nucleotide polymorphism in an ultra-conserved amino acid of the RING domain of uhrf1. (A–B) Live images of 5 dpf wildtype (A) and uhrf1b1115 mutant (B) larvae show that mutants have smaller eyes and heart edema compared to wildtype siblings. (C) Exon 16 of uhrf1 contains the mutation (red asterisk) in the b1115 allele. (D) Schematic of important protein domains of uhrf1 with a close-up of the seven conserved cysteines (Tauber and Fischle, 2015). (E) A non-synonymous single nucleotide polymorphism (T=>C) mutates the conserved cysteine 7 (red) to a Glycine. (F) Cysteine 7 (red) is conserved among phylogenetically diverse vertebrate species. (G) Complementation cross between the b1115 allele and the hi272 allele of uhrf1 shows failure to complement in 5 dpf larvae as demonstrated by fewer phox2b:EGFP enteric neurons (green; H) compared to wildtype (green; G). In both proximal (J) and distal (L) intestine, uhrf1b1115/hi272 mutants have fewer enteric neurons than wildtype siblings at 5 dpf (proximal I; distal K) as quantified in M and N. (M,N) Quantification of Elavl positive cells per 200 μm of proximal or distal intestine in wildtype or uhrf1b1115/hi272 mutant [wildtype n = 10 (proximal), n = 10 (distal); uhrf1b1115/hi272 n = 19 (proximal), n = 18 (distal)]. UBL, ubiquitin like; TTD, tandem tudor domain; PHD, plant homeodomain; SRA, SET and RING associated; PBR, polybasic region; RING, really interesting and new gene. A,B,G,H: side-views of whole-mount zebrafish larvae at 5 dpf. I–L: Confocal images of dissected intestines at 5 dpf. Scale bar = 100 μm in A,B,E,F; Scale bar = 50 μm in I-L. Unpaired t-test: **** ​= ​p ​< ​0.0001.