Bent DNA in the human adenovirus type 2 E1A enhancer is an architectural element for transcription stimulation

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Abstract

The upstream half of the human adenovirus type 2 enhancer adopts a curved DNA structure. Most of the enhancer elements are within the curvature, suggesting that this unusual structure is linked to enhancer function. To verify this experimentally, I constructed in vitro transcription assay systems which could distinguish any effects generated by conformational changes in a DNA template. The curved DNA conformation in the enhancer clearly affected the extent of the stimulation of the E1A gene transcription: assays using the wild-type DNA template showed that the moderately curved enhancer was superior to the highly curved enhancer in transcriptional stimulation. In additional experiments, the enhancer region was substituted with a curved DNA derived from the bacteriophage λ origin of replication. Assays using this mutant revealed that this curved segment could also act as an enhancer when it had the proper conformation. Consequently, DNA conformation may play a general role in transcriptional stimulation.

Original languageEnglish
Pages (from-to)27823-27828
Number of pages6
JournalJournal of Biological Chemistry
Volume271
Issue number44
Publication statusPublished - 1996
Externally publishedYes

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Human Adenoviruses
Transcription
Nucleic Acid Conformation
DNA
Conformations
Assays
Replication Origin
Bacteriophages
Genes

ASJC Scopus subject areas

  • Biochemistry

Cite this

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abstract = "The upstream half of the human adenovirus type 2 enhancer adopts a curved DNA structure. Most of the enhancer elements are within the curvature, suggesting that this unusual structure is linked to enhancer function. To verify this experimentally, I constructed in vitro transcription assay systems which could distinguish any effects generated by conformational changes in a DNA template. The curved DNA conformation in the enhancer clearly affected the extent of the stimulation of the E1A gene transcription: assays using the wild-type DNA template showed that the moderately curved enhancer was superior to the highly curved enhancer in transcriptional stimulation. In additional experiments, the enhancer region was substituted with a curved DNA derived from the bacteriophage λ origin of replication. Assays using this mutant revealed that this curved segment could also act as an enhancer when it had the proper conformation. Consequently, DNA conformation may play a general role in transcriptional stimulation.",
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