Fig. 1. Cloning of the human NUANCE gene. (A) The 21.8 kb cDNA was assembled from 13 overlapping clones obtained by RT- and RACE-PCR using mRNA from BL-60 cells as the template. (B) The exon-intron organization of the human NUANCE gene. The NUACE gene contains 114 protein-coding exons. Introns, represented by lines, are drawn to scale; exons are shown as vertical bars. Exon 1a, found in the EST BI026470, exon 8a, found in the short isoform NUANCE-ABD-S, exon 103a, found in the KIAA1011 protein (accession number AB023228), and the longer form of exon 104, which is found in DKFZp434H2235 mRNA (accession number AL117404), are represented by empty bars. The positions of Start and Stop codons are indicated by arrows. The annotated sequences of DKFZp434G173 (accession number NM_015180) and DKFZp434H2235 are underlined by solid lines. The estrogen receptor 2 gene (ESR2) located downstream is boxed. Alternative splicing at the 5' end (C) and at the 3' end (D) of the NUANCE gene. An additional exon 1a that encodes an alternative transcriptional start was found in EST BI026470. The boxes represent exons drawn to scale. Gray shading indicate coding regions; white boxes indicate untranslated regions. Aternatively spliced coding exons are shown in black. The locations of the ATG start codon and the TGA stop codon are indicated. The size of each exon and intron is given in Table 1.