The dual role of helix - loop - helix-zipper protein USF in ribosomal RNA gene transcription in vivo

Asish K. Ghosh, Prasun K. Datta, Samson T. Jacob*

*Corresponding author for this work

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

We have previously demonstrated that the core promoter of rat ribosomal RNA gene (rDNA) contains an E-box-like sequence to which the core promoter binding factor CPBF binds and that the 44 kDa subunit of this protein is inmunologically related to USF1, the helix - loop - helix-zipper DNA binding protein. Further, we showed that RNA polymerase I (pol I) transcription in vitro is competed by oligonucleotides containing USF-binding site, which suggested a key role for USF in rDNA transcription. To prove the potential role of USF in pol I transcription in vivo, USF1 and USF2 homodimers and USF1/USF2 heterodimer were overexpressed in CHO cells by transfection of the respective cDNAs. Co-transfection of a plasmid containing rDNA followed by primer extension analysis showed that overexpression of USF1 and USF2 as homodimers resulted in inhibition of rDNA transcription by as much as 85-90% whereas overexpression of USF1/USF2 in the heterodimeric form activated transcription approximately 3.5-fold. Transfection of mutant USF2 cDNA that is devoid of the basic DNA-binding domain produced only minimal inhibition of rDNA transcription. These data show that USF can modulate transcription of rRNA gene in vivo by functioning as a repressor (homodimer) or activator (heterodimer) of pol I transcription in vivo and suggest that inhibition of rDNA transcription may be responsible for the antiproliferative action of USF homodimers.

Original languageEnglish (US)
Pages (from-to)589-594
Number of pages6
JournalOncogene
Volume14
Issue number5
DOIs
StatePublished - Jan 1 1997

Fingerprint

Ribosomal DNA
rRNA Genes
Transfection
Proteins
Complementary DNA
Core Binding Factors
E-Box Elements
RNA Polymerase I
CHO Cells
Protein Subunits
DNA-Binding Proteins
Oligonucleotides
Plasmids
Binding Sites
DNA

Keywords

  • Activator
  • Repressor
  • Ribosomal RNA gene
  • Upstream stimulatory factor
  • pol I transcription

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

Cite this

Ghosh, Asish K. ; Datta, Prasun K. ; Jacob, Samson T. / The dual role of helix - loop - helix-zipper protein USF in ribosomal RNA gene transcription in vivo. In: Oncogene. 1997 ; Vol. 14, No. 5. pp. 589-594.
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The dual role of helix - loop - helix-zipper protein USF in ribosomal RNA gene transcription in vivo. / Ghosh, Asish K.; Datta, Prasun K.; Jacob, Samson T.

In: Oncogene, Vol. 14, No. 5, 01.01.1997, p. 589-594.

Research output: Contribution to journalArticle

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T1 - The dual role of helix - loop - helix-zipper protein USF in ribosomal RNA gene transcription in vivo

AU - Ghosh, Asish K.

AU - Datta, Prasun K.

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N2 - We have previously demonstrated that the core promoter of rat ribosomal RNA gene (rDNA) contains an E-box-like sequence to which the core promoter binding factor CPBF binds and that the 44 kDa subunit of this protein is inmunologically related to USF1, the helix - loop - helix-zipper DNA binding protein. Further, we showed that RNA polymerase I (pol I) transcription in vitro is competed by oligonucleotides containing USF-binding site, which suggested a key role for USF in rDNA transcription. To prove the potential role of USF in pol I transcription in vivo, USF1 and USF2 homodimers and USF1/USF2 heterodimer were overexpressed in CHO cells by transfection of the respective cDNAs. Co-transfection of a plasmid containing rDNA followed by primer extension analysis showed that overexpression of USF1 and USF2 as homodimers resulted in inhibition of rDNA transcription by as much as 85-90% whereas overexpression of USF1/USF2 in the heterodimeric form activated transcription approximately 3.5-fold. Transfection of mutant USF2 cDNA that is devoid of the basic DNA-binding domain produced only minimal inhibition of rDNA transcription. These data show that USF can modulate transcription of rRNA gene in vivo by functioning as a repressor (homodimer) or activator (heterodimer) of pol I transcription in vivo and suggest that inhibition of rDNA transcription may be responsible for the antiproliferative action of USF homodimers.

AB - We have previously demonstrated that the core promoter of rat ribosomal RNA gene (rDNA) contains an E-box-like sequence to which the core promoter binding factor CPBF binds and that the 44 kDa subunit of this protein is inmunologically related to USF1, the helix - loop - helix-zipper DNA binding protein. Further, we showed that RNA polymerase I (pol I) transcription in vitro is competed by oligonucleotides containing USF-binding site, which suggested a key role for USF in rDNA transcription. To prove the potential role of USF in pol I transcription in vivo, USF1 and USF2 homodimers and USF1/USF2 heterodimer were overexpressed in CHO cells by transfection of the respective cDNAs. Co-transfection of a plasmid containing rDNA followed by primer extension analysis showed that overexpression of USF1 and USF2 as homodimers resulted in inhibition of rDNA transcription by as much as 85-90% whereas overexpression of USF1/USF2 in the heterodimeric form activated transcription approximately 3.5-fold. Transfection of mutant USF2 cDNA that is devoid of the basic DNA-binding domain produced only minimal inhibition of rDNA transcription. These data show that USF can modulate transcription of rRNA gene in vivo by functioning as a repressor (homodimer) or activator (heterodimer) of pol I transcription in vivo and suggest that inhibition of rDNA transcription may be responsible for the antiproliferative action of USF homodimers.

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