Did nucleotides or amino acids drive evolutionary conservation of the WT1 ±KTS alternative splice?

Evolutionary comparisons frequently pinpoint crucial parts of a protein but, even within coding regions, nucleotides can do more than determine amino acid sequence. One highly conserved feature of the Wilms tumour suppressor gene, WT1, is the potential, following alternative pre-mRNA splicing, to insert three amino acids (KTS) between the third and fourth zinc fingers. The nucleotides at this position simultaneously define amino acids and the alternative splice site.

At the protein level this insertion influences DNA binding affinity and specificity, protein protein interactions and subnuclear localization. Mutations within the ±KTS splice junction lead to severe urogenital developmental abnormalities such as Frasier syndrome, indicating that the isoform ratio is critical for wild-type function. Using a series of site-directed mutations in both the genomic and cDNA context, the nucleotide amino acid relationship was investigated.

Mutational analysis within the cDNA suggests that the ... more.

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