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PMID: 7910795 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Deformed protein binding sites and cofactor binding sites are required for the function of a small segment-specific regulatory element in Drosophila embryos.

The EMBO journal ·Vol. 13 ·No. 10 ·1994-05-15 ·Pages 2362-77

Zeng C, Pinsonneault J, Gellon G, McGinnis N, McGinnis W

Abstract

How each of the homeotic selector proteins can regulate distinct sets of DNA target elements in embryos is not understood. Here we describe a detailed functional dissection of a small element that is specifically regulated by the Deformed homeotic protein. This 120 bp element (module E) is part of a larger 2.7 kb autoregulatory enhancer that maintains Deformed (Dfd) transcription in the epidermis of the maxillary and mandibular segments of Drosophila embryos. In vitro binding assays show that module E contains only one Dfd protein binding site. Mutations in the Dfd binding site that increase or decrease its in vitro affinity for Dfd protein generate parallel changes in the regulatory activity of module E in transgenic embryos, strong evidence that the in vitro-defined binding site is a direct target of Dfd protein in embryos. However, a monomer or multimer of the Dfd binding region alone is not sufficient to supply Dfd-dependent, segment-specific reporter gene expression. An analysis of a systematic series of clustered point mutations in module E revealed that an additional region containing an imperfect inverted repeat sequence is also required for the function of this homeotic protein response element. The Dfd binding site and the putative cofactor binding site(s) in the region of the inverted repeat are both necessary and in combination sufficient for the function of module E.

MeSH Terms
Animals Base Sequence DNA Mutational Analysis DNA-Binding Proteins/genetics,metabolism Drosophila Proteins Drosophila melanogaster/embryology,genetics Enhancer Elements, Genetic/genetics Gene Expression Regulation Genes, Homeobox Genes, Reporter Homeodomain Proteins Insect Hormones/genetics,metabolism Molecular Sequence Data Protein Binding Regulatory Sequences, Nucleic Acid/genetics Repetitive Sequences, Nucleic Acid Sequence Analysis, DNA Sequence Deletion Transcription, Genetic Transformation, Genetic
Chemicals
DNA-Binding Proteins Dfd protein, Drosophila Drosophila Proteins Homeodomain Proteins Insect Hormones
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Zeng C
Department of Molecular Biophysics, Yale University, New Haven, CT 06520-8114.
Pinsonneault J
Gellon G
McGinnis N
McGinnis W
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1994-05-15
Pages
2362-77
Language
English
Region
England
NLM ID
8208664
PMCID
PMC395101
Subset
IM
Grants
NICHD NIH HHS · HD28315 · United States
Databases
GENBANK
S69979
External Links
PubMed source PMC Full Text View journal details
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