DNA sequence determinants for binding of transformed Ah receptor to a dioxin-responsive enhancer

Eveline F. Yao, Michael S. Denison

Research output: Contribution to journalArticle

112 Citations (Scopus)

Abstract

We have utilized gel retardation analysis and DNA mutagenesis to examine the specific interaction of transformed guinea pig hepatic cytosolic TCDD·AhR complex with a dioxin-responsive element (DRE). Sequence alignment of the mouse CYPIA1 upstream DREs has identified a common invariant "core" consensus sequence of TNGCGTG flanked by several variable nucleotides. Competitive gel retardation analysis using a series of DRE oligonucleotides containing single or multiple base substitutions has allowed identification of those nucleotides important for TCDD·AhR·DRE complex formation. A putative TCDD·AhR DNA-binding consensus sequence of GCGTGNNA/TNNNC/G has been derived. The four core nucleotides, CGTG, appear to be critical for TCDD-inducible protein-DNA complex formation since their substitution decreased AhR binding affinity by 100-800-fold; the remaining conserved bases are also important, albeit to a lesser degree (3-5-fold). The 5′-ward thymine, present in the invariant core sequence of all the DREs identified to date, appears not to be involved in DNA binding of the AhR. The results obtained here indicate that although the primary interaction of the TCDD·AhR complex with the DRE occurs with the conserved "core" sequence, nucleotides flanking the core also contribute to the specificity of DRE binding.

Original languageEnglish (US)
Pages (from-to)5060-5067
Number of pages8
JournalBiochemistry
Volume31
Issue number21
StatePublished - 1992
Externally publishedYes

Fingerprint

Dioxins
DNA sequences
Nucleotides
DNA
Consensus Sequence
Substitution reactions
Gels
Mutagenesis
Thymine
Conserved Sequence
Sequence Alignment
Oligonucleotides
Guinea Pigs
Liver
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

DNA sequence determinants for binding of transformed Ah receptor to a dioxin-responsive enhancer. / Yao, Eveline F.; Denison, Michael S.

In: Biochemistry, Vol. 31, No. 21, 1992, p. 5060-5067.

Research output: Contribution to journalArticle

Yao, Eveline F. ; Denison, Michael S. / DNA sequence determinants for binding of transformed Ah receptor to a dioxin-responsive enhancer. In: Biochemistry. 1992 ; Vol. 31, No. 21. pp. 5060-5067.
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