DEV01611 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Mapping of the mol‾^/‾ mutation. Position of both the mol^tv53aand mol^st20 loci and foxa2 on a ZMAP panel of linkage group 17, showing both SSLP marker and gene positions. mol^tv53a embryos show 5/240 recombination events with z21703 and 8/68 with z21194. Radiaton hybrid and meiotic mapping places foxa2 in the same interval (Woods et al., 2000) (http://zfin.org/cgi-bin/webdriver?MIval=aa-markerview.apg&OID=ZDBGENE-980526-404). mol^tv53a embryos show 0/88 recombination events with a marker placed within foxa2 itself. Sequence analysis of mol^tv53a embryos identified a point mutation of C to T in nucleotide 496 of the open reading frame of foxa2, causing an amino acid change of glutamine 166 (CAA) to stop (TAA). The tv53a mutation is within the region encoding the forkhead domain of Foxa2 and causes a truncation (Fig. 1E) that is likely to abolish activity of the protein. Similarly, mol^st20 was placed on LG17, in proximity to both z21703 and z9830 (one recombinant in 20 meioses for each). Sequence analysis of two exons of foxa2 from fin clips of heterozygous adult carriers identified a point mutation of C to A, in nucleotide 534 of the open reading frame. This leads to an amino acid substitution of tyrosine 178 (TAC) to stop (TAA) (Fig. 1E). Similar to mol^tv53a, this results in a truncation of Foxa2 within the DNA binding forkhead domain. By scoring an Mse1 restriction enzyme polymorphism introduced by the C to A mutation, this lesion co-segregated with the mol^st20 mutation in 108 tested mutant embryos.