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Related Experiment Videos

Atlantic salmon HNF-3/forkhead: cDNA sequence, evolution, expression, and functional analysis.

C Stenson1, A McNair, L Byrnes

  • 1National Diagnostics Centre/BioResearch Ireland (NDC), Department of Microbiology, National University of Ireland, Galway.

DNA and Cell Biology
|February 11, 2000
PubMed
Summary
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Researchers isolated and characterized a novel Atlantic salmon HNF-3 gene, finding it lacks introns and is expressed early in development. This gene binds to specific DNA sequences, including a new site in the transferrin gene promoter.

Area of Science:

  • Molecular Biology
  • Developmental Biology
  • Genomics

Background:

  • Hepatocyte Nuclear Factor 3 (HNF-3) proteins are crucial transcription factors involved in development and cell differentiation.
  • Understanding HNF-3 homologs in non-model organisms like Atlantic salmon can provide insights into conserved regulatory mechanisms.

Purpose of the Study:

  • To isolate and characterize a cDNA encoding an HNF-3 family member from Atlantic salmon (Salmo salar L).
  • To investigate the expression pattern and functional properties of the identified Atlantic salmon HNF-3 (asHNF-3).
  • To identify novel binding sites for asHNF-3.

Main Methods:

  • cDNA isolation and sequencing.
  • Sequence alignment with known winged helix/forkhead proteins.

Related Experiment Videos

  • Comparison of cDNA and genomic DNA to detect introns.
  • Reverse Transcription Polymerase Chain Reaction (RT-PCR) for expression analysis in developmental stages and adult tissues.
  • Electrophoretic mobility shift assays (EMSAs) to assess DNA binding activity.

    • Main Results:

    • A cDNA encoding an Atlantic salmon HNF-3 (asHNF-3) was successfully isolated and characterized.
    • The asHNF-3 gene lacks introns, and its protein sequence shows high identity to other HNF-3 family members.
    • asHNF-3 is expressed in adult salmon liver, pancreas, and intestine, and is detectable before gastrulation during development.
    • Functional analysis confirmed that asHNF-3 can bind to HNF-3 consensus sequences.
    • A novel HNF3 binding site was identified in the promoter of the Atlantic salmon transferrin gene.

    Conclusions:

    • The study reports the first characterization of an HNF-3 family member in Atlantic salmon, revealing conserved structural and functional features.
    • The early expression and tissue distribution suggest a significant role for asHNF-3 in salmon development and physiology.
    • The identification of a novel binding site in the transferrin gene promoter opens avenues for further research into gene regulation in fish.