Analysis of core promoter sequences located downstream from the TATA element in the hsp70 promoter from Drosophila melanogaster

C. H. Wu, L. Madabusi, H. Nishioka, P. Emanuel, M. Sypes, I. Arkhipova, D. S. Gilmour

Research output: Contribution to journalArticle

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Abstract

TFIID recognizes multiple sequence elements in the hsp70 promoter of Drosophila. Here, we investigate the function of sequences downstream from the TATA element. A mutation in the initiator was identified that caused an eightfold reduction in binding of TFIID and a fourfold reduction in transcription in vitro. Another mutation in the +24 to +29 region was somewhat less inhibitory, but a mutation in the +14 to +19 region had essentially no effect. The normal promoter and the mutants in the initiator and the +24 to +29 region were transformed into flies by P element-mediated transformation. The initiator mutation reduced expression an average of twofold in adult flies, whereas the mutation in the +24 to +29 region had essentially no effect. In contrast, a promoter combining the two mutations was expressed an average of sixfold less than the wild type. The results suggest that the initiator and the +24 to +29 region could serve overlapping functions in vivo. Protein-DNA cross-linking was used to identify which subunits of TFIID contact the +24 to +29 region and the initiator. No specific subunits were found to cross-link to the +24 to +29 region. In contrast, the initiator cross-linked exclusively to dTAF230. Remarkably, dTAF230 cross-links approximately 10 times more efficiently to the nontranscribed strand than to the transcribed strand at the initiator.

Original languageEnglish (US)
Pages (from-to)1593-1602
Number of pages10
JournalMolecular and cellular biology
Volume21
Issue number5
DOIs
StatePublished - Feb 26 2001

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Drosophila melanogaster
Transcription Factor TFIID
Mutation
Diptera
Drosophila
DNA
Proteins

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Wu, C. H. ; Madabusi, L. ; Nishioka, H. ; Emanuel, P. ; Sypes, M. ; Arkhipova, I. ; Gilmour, D. S. / Analysis of core promoter sequences located downstream from the TATA element in the hsp70 promoter from Drosophila melanogaster. In: Molecular and cellular biology. 2001 ; Vol. 21, No. 5. pp. 1593-1602.
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Analysis of core promoter sequences located downstream from the TATA element in the hsp70 promoter from Drosophila melanogaster. / Wu, C. H.; Madabusi, L.; Nishioka, H.; Emanuel, P.; Sypes, M.; Arkhipova, I.; Gilmour, D. S.

In: Molecular and cellular biology, Vol. 21, No. 5, 26.02.2001, p. 1593-1602.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Analysis of core promoter sequences located downstream from the TATA element in the hsp70 promoter from Drosophila melanogaster

AU - Wu, C. H.

AU - Madabusi, L.

AU - Nishioka, H.

AU - Emanuel, P.

AU - Sypes, M.

AU - Arkhipova, I.

AU - Gilmour, D. S.

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