Position-dependent transcriptional regulation of the murine dihydrofolate reductase promoter by the E2F transactivation domain

Christopher J. Fry, Jill E. Slansky, Peggy J. Farnham

Research output: Contribution to journalArticle

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Abstract

Activity of the dihydrofolate reductase (dhfr) promoter increases at the G1-S-phase boundary of the cell cycle. Mutations that abolish protein binding to an E2F element in the dhfr promoter also abolish the G2-S-phase increase in dhfr transcription, indicating that transcriptional regulation is mediated by the E2F family of proteins. To investigate the mechanism by which E2F regulates dhfr transcription, we moved the E2F element upstream and downstream of its natural position in the promoter. We found that the E2F element confers growth regulation to the dhfr promoter only when it is proximal to the transcription start site. Using a heterologous E2F element, we showed that position-dependent regulation is a property that is promoter specific, not E2F element specific. We demonstrated that E2F-mediated growth regulation of dhfr transcription requires activation of the dhfr promoter in S phase and that the C-terminal activation domains of E2F1, E2F4, and E2F5, when fused to the Gal4 DNA binding domain, are sufficient to specify position-dependent activation. To further investigate the role of activation in dhfr regulation, we tested other transactivation domains for their ability to activate the dhfr promoter. We found that the N-terminal transactivation domain of VP16 cannot activate the dhfr promoter. We propose that, unlike other E2F-regulated promoters, robust transcription from the dhfr promoter requires an E2F transactivation domain close to the transcription start site.

Original languageEnglish (US)
Pages (from-to)1966-1976
Number of pages11
JournalMolecular and Cellular Biology
Volume17
Issue number4
StatePublished - Apr 1997
Externally publishedYes

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Tetrahydrofolate Dehydrogenase
Transcriptional Activation
S Phase
Transcription Initiation Site
E2F Transcription Factors
G2 Phase
G1 Phase
Growth
Protein Binding
Cell Cycle

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Position-dependent transcriptional regulation of the murine dihydrofolate reductase promoter by the E2F transactivation domain. / Fry, Christopher J.; Slansky, Jill E.; Farnham, Peggy J.

In: Molecular and Cellular Biology, Vol. 17, No. 4, 04.1997, p. 1966-1976.

Research output: Contribution to journalArticle

Fry, Christopher J. ; Slansky, Jill E. ; Farnham, Peggy J. / Position-dependent transcriptional regulation of the murine dihydrofolate reductase promoter by the E2F transactivation domain. In: Molecular and Cellular Biology. 1997 ; Vol. 17, No. 4. pp. 1966-1976.
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