Protein-DNA interactions at the H4-site III upstream transcriptional element of a cell cycle regulated histone H4 gene: Differences in normal versus tumor cells

C. Willemien van der Houven van Oordt, Andre J van Wijnen, R. Carter, K. Soprano, J. B. Lian, G. S. Stein, J. L. Stein

Research output: Contribution to journalArticle

11 Scopus citations


Upstream sequences of the H4 histone gene FO108 located between nt -418 to -213 are stimulatory for in vivo transcription. This domain contains one protein/DNA interaction site (H4-Site III) that binds factor H4UA-1. Based on methylation interference, copper-phenanthroline protection, and competition assays, we show that H4UA-1 interacts with sequences between nt -345 to -332 containing an element displaying sequence-similarity with the thyroid hormone response element (TRE). Using gel retardation assays, we also demonstrate that H4UA-1 binding activity is abolished at low concentrations of Zn2+ (0.75 mM), a characteristic shared with the thyroid hormone (TH) receptor DNA binding protein. Interestingly, phosphatase-treatment of nuclear proteins inhibit formation of the H4UA-1 protein/DNA complex, although a complex with higher mobility (H4UA-1b) can be detected; both complexes share identical protein-DNA contacts and competition behaviors. These findings suggest that phosphorylation may be involved in the regulation of H4-Site III protein/DNA interactions by directly altering protein/protein associations. H4-Site III interactions were examined in several cell culture systems during cell growth and differentiation. We find that H4UA-1 binding activity is present during the cell cycle of both normal diploid and transformed cells. However, during differentiation of normal diploid rat calvarial osteoblasts, we observe a selective loss of the H4UA-1/H4-Site III interaction, concomitant with an increase of the H4UA-1b/H4-Site III complex, indicating modifications in the heteromeric nature of protein/DNA interactions during downregulation of transcription at the cessation of proliferation. Transformed cells have elevated levels of H4UA-1, whereas H4UA-1b is predominantly present in normal diploid cells; this alteration in the ratio of H4UA-1 and H4UA-1b binding activities may reflect deregulation of H4-Site III interactions in transformed cells. We propose that H4-Site III interactions may contribute, together with protein/DNA interactions at proximal regulatory sequences, in determining the level of H4-FO108 histone gene transcription.

Original languageEnglish (US)
Pages (from-to)93-110
Number of pages18
JournalJournal of Cellular Biochemistry
Issue number1
StatePublished - 1992
Externally publishedYes



  • Cell growth
  • Differentiation
  • Distal promoter elements
  • DNA-binding proteins
  • Proliferation

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Willemien van der Houven van Oordt, C., van Wijnen, A. J., Carter, R., Soprano, K., Lian, J. B., Stein, G. S., & Stein, J. L. (1992). Protein-DNA interactions at the H4-site III upstream transcriptional element of a cell cycle regulated histone H4 gene: Differences in normal versus tumor cells. Journal of Cellular Biochemistry, 49(1), 93-110.