Insensitivity of the present hsp26 chromatin structure to a TATA box mutation in Drosophila

Quinn Lu; Lori L Wallrath; Peter A Emanuel; Sarah C.R Elgin; David S Gilmour

doi:10.1016/S0021-9258(17)40766-6

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Insensitivity of the present hsp26 chromatin structure to a TATA box mutation in Drosophila

Journal article

Open access

Peer reviewed

Insensitivity of the present hsp26 chromatin structure to a TATA box mutation in Drosophila

Quinn Lu, Lori L Wallrath, Peter A Emanuel, Sarah C.R Elgin and David S Gilmour

The Journal of biological chemistry, Vol.269(22), pp.15906-15911

06/03/1994

DOI: 10.1016/S0021-9258(17)40766-6

PMID: 8195245

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Abstract

The role of the TATA element in establishing the chromatin structure and inducible transcription of the Drosophila melanogaster hsp26 gene has been analyzed. An hsp26/lacZ fusion gene with a mutant promoter, in which the TATA box sequence TATAAA was changed to CCCAAA, was introduced into Drosophila by P-element transformation. The mutation had little effect on formation of the preset chromatin structure observed prior to induction. However, the mutation dramatically reduced transcription levels following heat shock. Northern analysis indicated that weak, inducible expression of the mutant promoter occurred within the same period of heat shock as for the normal promoter, suggesting that TFIID was associated with the mutant promoter prior to heat shock. Biochemical analysis showed that the mutant promoter still bound TFIID in vitro, but with 3-5-fold less affinity than the normal promoter. DNase I footprinting revealed that the conformation of the TFIID-DNA complex differed significantly from that of the normal promoter. These results indicate that alterations in the conformation or the stability of the TFIID-DNA complex drastically reduce the level of induction, but do not dramatically affect chromatin structure formation. Formation of the requisite chromatin structure is either independent of, or highly tolerant of, changes in the TFIID-DNA complex.

Gene Expression

Animals

Animals, Genetically Modified

Base Sequence

beta-Galactosidase - biosynthesis

beta-Galactosidase - genetics

Blotting, Northern

Chromatin - chemistry

Chromatin - metabolism

DNA - metabolism

Drosophila melanogaster - genetics

Drosophila melanogaster - metabolism

Drosophila Proteins

Heat-Shock Proteins - biosynthesis

Heat-Shock Proteins - genetics

Kinetics

Molecular Sequence Data

Mutagenesis, Site-Directed

Oligodeoxyribonucleotides

Restriction Mapping

TATA Box

Transcription Factor TFIID

Transcription Factors - metabolism

Transcription, Genetic

Details

Title: Subtitle: Insensitivity of the present hsp26 chromatin structure to a TATA box mutation in Drosophila
Creators: Quinn Lu - Department of Biology, Washington University, St. Louis, Missouri 63130
Lori L Wallrath - Washington University in St. Louis
Peter A Emanuel - Washington University in St. Louis
Sarah C.R Elgin - Washington University in St. Louis
David S Gilmour - Washington University in St. Louis
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.269(22), pp.15906-15911
DOI: 10.1016/S0021-9258(17)40766-6
PMID: 8195245
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Grant note: GM 31532 / NIGMS NIH HHS GM47477 / NIGMS NIH HHS GM14516 / NIGMS NIH HHS
Language: English
Date published: 06/03/1994
Academic Unit: Biochemistry and Molecular Biology; University College Courses
Record Identifier: 9984293079002771

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