A POU homeo domain protein related to dPOU-19/pdm-1 binds to the regulatory DNA necessary for vital expression of the Drosophila choline acetyltransferase gene

T Kitamoto; P M Salvaterra

doi:10.1523/JNEUROSCI.15-05-03509.1995

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A POU homeo domain protein related to dPOU-19/pdm-1 binds to the regulatory DNA necessary for vital expression of the Drosophila choline acetyltransferase gene

Journal article

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Peer reviewed

A POU homeo domain protein related to dPOU-19/pdm-1 binds to the regulatory DNA necessary for vital expression of the Drosophila choline acetyltransferase gene

T Kitamoto and P M Salvaterra

The Journal of neuroscience, Vol.15(5 Pt 1), pp.3509-3518

05/1995

DOI: 10.1523/JNEUROSCI.15-05-03509.1995

PMCID: PMC6578218

PMID: 7751926

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Abstract

Expression of the choline acetyltransferase (ChAT) gene in Drosophila melanogaster is responsible for production of the neurotransmitter acetylcholine and is necessary for viability. In previous studies, we have shown that the regulatory region for normal ChAT expression is large and composed of multiple regulatory elements (Kitamoto et al., 1992; Kitamoto and Salvaterra, 1993). In this study, using various lengths of 5' flanking DNA fused to wild type ChAT cDNA, we have defined a 0.3 kilobase (kb) region of the cis-regulatory DNA, which is essential for restoring viability of Cha lethal mutants. DNase I footprinting analysis of this 0.3 kb DNA revealed a protected 22 bp sequence that contains an octamer-like motif (ATTCAAAT) with one base difference from the consensus octamer motif (ATGCAAAT). Electrophoretic mobility shift assays and Southwestern blot analysis confirmed the presence of specific binding factor(s) for the 22 bp sequence in embryo nuclear extracts, and competition studies established the importance of the octamer-like motif for high-affinity binding. Using the 22-mer as a probe, we have isolated a cDNA clone encoding the Drosophila POU homeo domain protein, dPOU-19/pdm-1, whose target genes and specific binding sequences have not been identified. We propose that vital expression of the Drosophila ChAT gene is regulated by a member of the dPOU-19/pdm-1 putative transcription factor family.

Consensus Sequence

Homeodomain Proteins - metabolism

Regulatory Sequences, Nucleic Acid

Molecular Sequence Data

Immunoblotting

Drosophila melanogaster - genetics

Choline O-Acetyltransferase - biosynthesis

DNA-Binding Proteins - metabolism

Genes, Lethal

Base Sequence

Binding Sites

Genetic Linkage

Genes, Insect

Transformation, Genetic

Drosophila Proteins

POU Domain Factors

Animals, Genetically Modified

Chromosome Mapping

DNA - metabolism

Gene Expression Regulation, Enzymologic

Phenotype

Animals

beta-Galactosidase - biosynthesis

Mutagenesis

Embryo, Nonmammalian

Drosophila melanogaster - enzymology

Transcription Factors

Details

Title: Subtitle: A POU homeo domain protein related to dPOU-19/pdm-1 binds to the regulatory DNA necessary for vital expression of the Drosophila choline acetyltransferase gene
Creators: T Kitamoto - Division of Neurosciences, Institute of the City of Hope, Duarte, California 91010, USA
P M Salvaterra
Resource Type: Journal article
Publication Details: The Journal of neuroscience, Vol.15(5 Pt 1), pp.3509-3518
Publisher: United States
DOI: 10.1523/JNEUROSCI.15-05-03509.1995
PMID: 7751926
PMCID: PMC6578218
ISSN: 0270-6474
eISSN: 1529-2401
Language: English
Date published: 05/1995
Academic Unit: Iowa Neuroscience Institute; Anesthesia; Neuroscience and Pharmacology
Record Identifier: 9984006403702771

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