An AP-1 binding site in the upstream region of DEB-A is part of a developmentally regulated negative element

G. L. Wang, Elliott S. Goldstein

Research output: Contribution to journalArticle

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Abstract

The Deb-A gene from Drosophila melanogaster encodes a small membrane-associated protein, regulated during development, with peak abundance at 12-15 h of embryogenesis. The cis-acting regulatory elements that control expression of Deb-A during embryogenesis were localized using a somatic transformation assay. The Adh gene of D. melanogaster was used as a 'reporter' gene. The promoterless ADH coding sequence was fused to the 5′-upstream control region of Deb-A. Deletions were introduced into the 5′-region using various restriction sites and Bal31 deletion mutagenesis. A negative regulatory element, or silencer, was localized to a segment 47 base pairs long, between -395 and -442. It is responsible for 80% of the repression of gene expression during late development and reduces levels of Deb-A RNA nearly 5-fold. This negative element is temporally functional. It becomes active after 15 h of embryogenesis and it has no effect on gene expression prior to that. Within this negative element of 47 base pairs, two footprint regions were protected from DNase I digestion by embryonic nuclear extracts: one region contains an AP-1 binding site, but the other footprint is due to unknown element(s). High molecular weight DNA-protein complexes on an oligonucleotide probe spanning the AP-1 binding site were identified in gel retardation assays using partially purified bacterially expressed Djun protein or nuclear extracts from Drosophila embryos. These data suggest that the AP-1 site may be partly responsible for decreasing Deb-A expression during the late embryonic developmental stages of D. melanogaster.

Original languageEnglish (US)
Pages (from-to)94-104
Number of pages11
JournalBBA - Gene Structure and Expression
Volume1216
Issue number1
DOIs
StatePublished - Oct 19 1993

Fingerprint

Transcription Factor AP-1
Drosophila melanogaster
Embryonic Development
Genes
Binding Sites
Gene expression
Base Pairing
Assays
Transcriptional Silencer Elements
Gene Expression
Mutagenesis
Oligonucleotide Probes
Deoxyribonuclease I
Electrophoretic Mobility Shift Assay
Nuclear Proteins
Reporter Genes
Drosophila
Digestion
Membrane Proteins
Proteins

Keywords

  • (Drosophila)
  • AP-1
  • Deb-A gene
  • jun proto-oncogene

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Genetics
  • Structural Biology

Cite this

An AP-1 binding site in the upstream region of DEB-A is part of a developmentally regulated negative element. / Wang, G. L.; Goldstein, Elliott S.

In: BBA - Gene Structure and Expression, Vol. 1216, No. 1, 19.10.1993, p. 94-104.

Research output: Contribution to journalArticle

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