Multipartite structure of a negative regulatory element associated with a steroid hormone-inducible promoter

Jae Lee, Patricia G. Moffitt, Kimberlin L. Morley, David O. Peterson

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The steroid hormone-inducible promoter from mouse mammary tumor virus is associated with a distal negative regulatory element that represses its inherent basal activity. Deletion analysis localized the sequences required for repression to 64 base pairs of DNA between -427 and -364 with respect to the transcription initiation site. Transient transfection experiments with a series of linker scanning and small internal deletion mutations revealed two mutation-sensitive domains separated by a region of relative resistance to sequence alterations. DNase I footprinting and gel electrophoresis mobility shift experiments with crude nuclear extracts identified at least one protein-binding site within each of the two mutation-sensitive regions. An oligonucleotide corresponding to one of these sites is able to repress transcription, but only when linked to the promoter in multiple copies. This negative regulatory element functions synergistically with a promoter proximal negative element to mediate efficient promoter repression, selectively affecting basal relative to steroid hormone-induced transcription and thus increasing the ratio of promoter activity observed in the presence and absence of hormone.

Original languageEnglish (US)
Pages (from-to)24101-24108
Number of pages8
JournalJournal of Biological Chemistry
Volume266
Issue number35
StatePublished - Dec 15 1991
Externally publishedYes

Fingerprint

Steroid hormones
Transcription
Steroids
Hormones
Transcription Initiation Site
Deoxyribonuclease I
Electrophoresis
Viruses
Oligonucleotides
Mouse mammary tumor virus
Tumors
Mutation
Sequence Deletion
Gels
Experiments
Binding Sites
Complex Mixtures
Scanning
Protein Binding
Base Pairing

ASJC Scopus subject areas

  • Biochemistry

Cite this

Multipartite structure of a negative regulatory element associated with a steroid hormone-inducible promoter. / Lee, Jae; Moffitt, Patricia G.; Morley, Kimberlin L.; Peterson, David O.

In: Journal of Biological Chemistry, Vol. 266, No. 35, 15.12.1991, p. 24101-24108.

Research output: Contribution to journalArticle

Lee, Jae ; Moffitt, Patricia G. ; Morley, Kimberlin L. ; Peterson, David O. / Multipartite structure of a negative regulatory element associated with a steroid hormone-inducible promoter. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 35. pp. 24101-24108.
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