Sp1 mediates cell proliferation-dependent regulation of rat DNA topoisomerase IIα gene promoter

Jeong Ho Yoon, Jeong Kee Kim, Geun Bae Rha, Misook Oh, Se Ho Park, Rho Hyun Seong, Seung Hwan Hong, Sang Dai Park

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

DNA topoisomerase IIα (topo IIα) is an essential nuclear enzyme required for chromosome segregation during mitosis. Consistent with its critical role in cell division is the fact that the expression of the gene for topo IIα is strongly regulated by the proliferation state of cells. Using a transient expression system, we determined the contribution of putative cis-acting elements in its promoter region to its basal level and cell proliferation-dependent transcription. Experiments with 5' and/or 3' serial deletion and site-directed mutation revealed that (1) maximal promoter activity resides in the fragment extending to position -663 bp from the ATG initiation codon, (2) minimal promoter activity is harboured at -195 bp, (3) the defined minimal promoter contains only two putative elements, inverted CCAAT box 4 (ICB4) (-166 to -162 bp) and the most proximal GC-rich box in the promoter (GC2) (-149 to -143 bp), and (4) ICB4 is most important in the basal-level transcription of the gene for rat topo IIα. The luciferase activities of the mutated reporter plasmids in G(O)-arrested and exponentially growing cells showed that proliferation-specific regulation is controlled mainly by GC2. Electrophoretic mobility-shift assays indicated that Sp1 binds specifically to the GC2 site. The extent of DNA-protein complex formation increases after the stimulation of cells to proliferate. These results indicate that the increased binding activity of Sp1 to GC2 is important in the up-regulation of the gene for topo IIα in growing cells.

Original languageEnglish
Pages (from-to)367-374
Number of pages8
JournalBiochemical Journal
Volume344
Issue number2
DOIs
Publication statusPublished - 1999 Dec 1

Keywords

  • Basal-level transcription
  • Upregulation
  • cis-acting elements

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Yoon, J. H., Kim, J. K., Rha, G. B., Oh, M., Park, S. H., Seong, R. H., Hong, S. H., & Park, S. D. (1999). Sp1 mediates cell proliferation-dependent regulation of rat DNA topoisomerase IIα gene promoter. Biochemical Journal, 344(2), 367-374. https://doi.org/10.1042/0264-6021:3440367