Direct interaction of the human cytomegalovirus IE86 protein with the cis repression signal does not preclude TBP from binding to the TATA box

Ray Jupp, Stefan Hoffmann, Alison Depto, Richard M. Stenberg, Peter Ghazal, Jay Nelson

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

The human cytomegalovirus major immediate-early gene encodes several protein isoforms which autoregulate the major immediate-early promoter (MIEP). One of these isoforms, the IE86 protein, represses the MIEP through a DNA sequence located between the TATA box and the transcription initiation site, designated the cis repression signal (crs). Through mutational analysis, amino acid domains within IE86 responsible for binding the crs element were located at the C terminus. Mutation of the putative zinc finger domain, which precluded IE86 from binding DNA, converted the protein from a repressor of MIEP transcription into an activator. DNase I protection analysis demonstrated that the IE86 footprint overlapped the sequence protected by the TATA-binding protein (TBP). Investigation of whether IE86 was able to displace TBP from DNA revealed that both proteins could bind DNA simultaneously. However, higher concentrations of IE86 were required to obtain protection of the crs element in the presence of prebound TBP. Similarly, higher concentrations of TBP were required to obtain protection in the presence of prebound IE86. These observations indicate that steric hinderance impairs but does not prevent both proteins from binding DNA synchronously.

Original languageEnglish (US)
Pages (from-to)5595-5604
Number of pages10
JournalJournal of Virology
Volume67
Issue number9
StatePublished - Sep 1993

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Human herpesvirus 5
TATA-Box Binding Protein
TATA box
TATA Box
Protein Binding
binding proteins
protein isoforms
promoter regions
DNA-Binding Proteins
Protein Isoforms
DNA
proteins
transcription (genetics)
deoxyribonuclease I
Immediate-Early Genes
DNA-binding proteins
Transcription Initiation Site
Deoxyribonuclease I
zinc finger motif
Zinc Fingers

ASJC Scopus subject areas

  • Immunology

Cite this

Direct interaction of the human cytomegalovirus IE86 protein with the cis repression signal does not preclude TBP from binding to the TATA box. / Jupp, Ray; Hoffmann, Stefan; Depto, Alison; Stenberg, Richard M.; Ghazal, Peter; Nelson, Jay.

In: Journal of Virology, Vol. 67, No. 9, 09.1993, p. 5595-5604.

Research output: Contribution to journalArticle

Jupp, Ray ; Hoffmann, Stefan ; Depto, Alison ; Stenberg, Richard M. ; Ghazal, Peter ; Nelson, Jay. / Direct interaction of the human cytomegalovirus IE86 protein with the cis repression signal does not preclude TBP from binding to the TATA box. In: Journal of Virology. 1993 ; Vol. 67, No. 9. pp. 5595-5604.
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