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Microarray analysis detects novel Pax3 downstream target genes.

C S Mayanil1, D George, L Freilich

  • 1Pediatric Brain Tumor Research Program, Department of Neurological Surgery, Northwestern University Medical School and Children's Memorial Institute of Education and Research, Chicago, Illinois 60614, USA.

The Journal of Biological Chemistry
|October 9, 2001
PubMed
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Paired box 3 (Pax3) transcription factor regulates gene expression. This study identified new downstream targets of Pax3, including alpha-2,8-polysialyltransferase (STX), crucial for neural development.

Area of Science:

  • Developmental Biology
  • Molecular Biology
  • Genetics

Background:

  • Pax3 is a crucial transcription factor for embryonic development, including neural tube and neural crest formation.
  • Previous research indicated Pax3's role in up-regulating alpha-2,8-polysialyltransferase (STX) gene expression and polysialic acid levels.

Purpose of the Study:

  • To identify novel downstream target genes of the Pax3 transcription factor.
  • To further investigate the regulatory relationship between Pax3 and STX.

Main Methods:

  • Utilized oligonucleotide arrays and RNA from stable Pax3 transfectants to screen for gene expression changes.
  • Confirmed array data using cDNA expression arrays and real-time quantitative polymerase chain reaction.
  • Employed promoter-luciferase reporter assays and electromobility shift assays to validate STX as a direct target.

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Main Results:

  • A total of 270 genes showed altered expression in Pax3 transfectants compared to controls.
  • Several identified genes possess potential Pax3 binding motifs in their promoter regions.
  • Confirmed STX as a direct downstream target of Pax3 through functional assays.

Conclusions:

  • Pax3 regulates a significant number of genes involved in cellular functions.
  • STX is a validated downstream target of Pax3, expanding the understanding of its regulatory network.
  • Several novel candidate genes are proposed as potential downstream targets of Pax3 for future research.