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Related Experiment Videos

c-myc activates RCC1 gene expression through E-box elements

M Tsuneoka1, F Nakano, H Ohgusu

  • 1Institute of Life Science, Kurume University, Fukuoka, Japan.

Oncogene
|May 15, 1997
PubMed
Summary

Proto-oncogene c-myc regulates mammalian cell proliferation by controlling RCC1 gene expression. This study demonstrates c-Myc directly binds to E-box elements in the RCC1 gene, confirming its role in cell growth.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Proto-oncogene c-myc is crucial for mammalian cell proliferation.
  • c-Myc functions as a transcription factor, but its regulated genes and precise role remain unclear.
  • RCC1 (Regulator of Chromosome Condensation 1) is essential for cell proliferation.

Purpose of the Study:

  • To investigate the relationship between c-myc expression and RCC1 gene expression.
  • To determine if c-Myc directly regulates RCC1 gene expression.
  • To identify the specific regulatory elements involved in c-myc-mediated RCC1 expression.

Main Methods:

  • Overexpression of c-myc in Rat 3Y1 cells and analysis of RCC1 mRNA levels.
  • Use of a chimeric c-myc protein to study its effect on RCC1 mRNA expression.

Related Experiment Videos

  • Construction of a reporter plasmid linking RCC1 gene fragments to luciferase cDNA for transient expression assays in HeLa cells.
  • Site-directed mutagenesis of E-box elements (CACGTG) in the reporter plasmid.
  • Gel retardation assays to assess c-Myc/Max binding to RCC1 gene sequences.

    • Main Results:

    • Serum stimulation increased both c-myc and RCC1 expression in HeLa cells.
    • Overexpression of c-myc led to a twofold increase in RCC1 mRNA levels.
    • Activation of a chimeric c-myc protein resulted in a twofold increase in RCC1 mRNA.
    • Co-transfection with c-myc dose-dependently stimulated luciferase activity driven by RCC1 gene fragments up to 2.5-fold.
    • Destruction of E-box elements abolished c-myc-mediated stimulation of reporter gene activity.
    • Gel retardation assays indicated that c-Myc/Max complex binds to RCC1 E-box elements in vitro.

    Conclusions:

    • c-Myc directly regulates the expression of the RCC1 gene.
    • Regulation occurs through specific E-box (CACGTG) elements located downstream of the RCC1 transcription start site.
    • These findings elucidate a novel mechanism by which c-Myc contributes to mammalian cell proliferation.