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Related Experiment Videos

Molecular mechanisms underlying SHP-1 gene expression.

Hing Wo Tsui1, Kathleen Hasselblatt, Alberto Martin

  • 1Division of Cellular & Molecular Biology, Toronto Western Research Institute, University Health Network, Toronto, Ontario, Canada.

European Journal of Biochemistry
|June 20, 2002
PubMed
Summary

This study reveals that upstream stimulatory factors (USFs) regulate the expression of SHP-1, a protein-tyrosine phosphatase. USFs, which have antiproliferative functions, interact with the SHP-1 promoter, influencing its activity.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • SHP-1 (protein-tyrosine phosphatase) is a negative regulator in signaling pathways, predominantly in hematopoietic and epithelial cells.
  • Two promoters and two transcript types (I and II) regulate SHP-1 expression, with identical cDNA sequences except for the 5' untranslated and initial coding regions.

Purpose of the Study:

  • To investigate promoter usage of SHP-1 in human and murine cells.
  • To identify regulatory elements and factors involved in SHP-1 gene expression, particularly in epithelial cells.
  • To explore the role of USFs in SHP-1 regulation.

Main Methods:

  • Comparative analysis of SHP-1 promoter usage in human and murine hematopoietic and epithelial cells.
  • Investigation of 4beta-phorbol 12-myristate 13-acetate's effect on human (I)SHP-1 transcript expression in SKOV3 cells.

Related Experiment Videos

  • Deletion analysis of a 12-bp repeat in the distal SHP-1 promoter.
  • Electrophoretic mobility shift assays (EMSAs) and supershift experiments to identify protein interactions with the promoter repeat.

    • Main Results:

    • Promoter usage for SHP-1 is similar in mouse and human hematopoietic cells but differs in epithelial cells, where only (I)SHP-1 transcripts are expressed.
    • 4beta-phorbol 12-myristate 13-acetate up-regulates human (I)SHP-1 transcript expression in SKOV3 cells, with potential involvement of nuclear factor-kappaB.
    • Deletion of one copy of a 12-bp repeat in the distal promoter significantly decreased promoter activity.
    • USF1 and USF2 were shown to hetero-dimerize and bind to this 12-bp repeat in the SHP-1 promoter.

    Conclusions:

    • USFs may regulate SHP-1 expression through interaction with a specific promoter element.
    • This regulation is significant given SHP-1's role as a negative growth regulator and USFs' antiproliferative functions.
    • Differential promoter usage in epithelial cells highlights cell-type-specific regulation of SHP-1.