Alternative splicing of Wilms tumor suppressor 1 (Wt1) exon 4 results in protein isoforms with different functions

Dev Biol. 2014 Sep 1;393(1):24-32. doi: 10.1016/j.ydbio.2014.06.026. Epub 2014 Jul 8.

Abstract

The Wilms tumor suppressor gene Wt1 encodes a zinc finger transcription factor that is essential for development of multiple organs including kidneys, gonads, spleen and heart. In mammals Wt1 comprises 10 exons with two characteristic splicing events: inclusion or skipping of exon 5 and alternative usage of two splice donor sites between exons 9 and 10. Most fish including zebrafish and medaka possess two wt1 paralogs, wt1a and wt1b, both lacking exon 5. Here we have characterized wt1 in guppy, platyfish and the short-lived African killifish Nothobranchius furzeri. All fish except zebrafish show alternative splicing of exon 4 of wt1a but not of wt1b with the wt1a(-exon 4) isoform being the predominant splice variant. With regard to function, Wt1a(+exon 4) showed less dimerization but stimulated transcription more effectively than the Wt1a(-exon 4) isoform. A specific knockdown of wt1a exon 4 in zebrafish was associated with anomalies in kidney development demonstrating a physiological function for Wt1a exon 4. Interestingly, alternative splicing of exon 4 seems to be an early evolutionary event as it is observed in the single wt1 gene of the sturgeon, a species that has not gone through teleost-specific genome duplication.

Keywords: Glomerulus; Kidney development; Morpholino; Pronephros; Transcription factor; Zebrafish.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing / genetics*
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • Cyprinodontiformes / embryology
  • Cyprinodontiformes / genetics
  • Fishes / abnormalities*
  • Fishes / genetics
  • Fundulidae / embryology
  • Fundulidae / genetics
  • Gene Expression Regulation, Developmental
  • Gene Knockout Techniques
  • Kidney / abnormalities*
  • Molecular Sequence Data
  • Morpholinos / genetics
  • Oryzias / embryology
  • Oryzias / genetics
  • Poecilia / embryology
  • Poecilia / genetics
  • Pronephros / abnormalities*
  • Protein Isoforms / genetics
  • Protein Multimerization
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Two-Hybrid System Techniques
  • WT1 Proteins / genetics*
  • Zebrafish / embryology
  • Zebrafish / genetics

Substances

  • Morpholinos
  • Protein Isoforms
  • WT1 Proteins

Associated data

  • GENBANK/KF601247
  • GENBANK/KF601248
  • GENBANK/KF601249
  • GENBANK/KF601250
  • GENBANK/KF601251
  • GENBANK/KF601252
  • GENBANK/KF601253
  • GENBANK/KF601254