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We used the reverse transcription-polymerase chain reaction (RT-PCR) to amplify choline acetyltransferase (ChAT) mRNA fragments from two temperature-sensitive alleles of Drosophila melanogaster, Cha(ts1) and Cha(ts2). Single base substitutions in the mutants (T1614A in Cha(ts1) and G1596A in Cha(ts2)) would result in amino acid changes for ChAT protein (Met403Lys in Ch(ts1) and Arg397His in Cha(ts2)). These base substitutions were confirmed in mRNA extracted from homozygous mutants using a Single Nucleotide Primer Extension assay (SNuPE) and are sufficient to produce thermolabile enzyme. Our results indicate that these temperature-sensitive mutants are point mutations in the structural gene for ChAT. Using a quantitative SNuPE assay we also show that similar levels of Cha(ts) and wild type transcripts are present in heterozygous flies (Cha(ts1)/+ and Cha(Ts2)/+) at both restrictive and permissive temperatures. This contrasts with RNase protection assays of ChAT mRNA in homozygous mutant animals where the levels of mutant mRNA decrease at restrictive temperature.
Bartus,
The cholinergic hypothesis of geriatric memory dysfunction.
1982, Pubmed
Bartus,
The cholinergic hypothesis of geriatric memory dysfunction.
1982,
Pubmed
Carbini,
Functional analysis of conserved histidines in choline acetyltransferase by site-directed mutagenesis.
1993,
Pubmed
Davies,
The neurochemistry of Alzheimer's disease and senile dementia.
1983,
Pubmed
Gorczyca,
Identification of a cholinergic synapse in the giant fiber pathway of Drosophila using conditional mutations of acetylcholine synthesis.
1984,
Pubmed
Greenwood,
Single nucleotide primer extension: quantitative range, variability, and multiplex analysis.
1996,
Pubmed
Itoh,
Cloning of Drosophila choline acetyltransferase cDNA.
1986,
Pubmed
Kitamoto,
Structure and organization of the Drosophila cholinergic locus.
1998,
Pubmed
McCaman,
Single RNA species injected in Xenopus oocyte directs the synthesis of active choline acetyltransferase.
1988,
Pubmed
,
Xenbase
Salvaterra,
Regulation of choline acetyltransferase.
1989,
Pubmed
Salvaterra,
Choline acetyltransferase and acetylcholine levels in Drosophila melanogaster: a study using two temperature-sensitive mutants.
1985,
Pubmed
Singer-Sam,
A sensitive, quantitative assay for measurement of allele-specific transcripts differing by a single nucleotide.
1992,
Pubmed
Sugihara,
Genomic organization of Drosophila choline acetyltransferase.
1991,
Pubmed
Sugihara,
Drosophila choline acetyltransferase uses a non-AUG initiation codon and full length RNA is inefficiently translated.
1990,
Pubmed
Tajima,
Sequence of choline acetyltransferase temperature-sensitive mutants determined by the polymerase chain reaction.
1990,
Pubmed
Takagawa,
Analysis of choline acetyltransferase protein in temperature sensitive mutant flies using newly generated monoclonal antibody.
1996,
Pubmed
Wu,
Choline acetyltransferase: celebrating its fiftieth year.
1994,
Pubmed
Yasuyama,
Differential regulation of choline acetyltransferase expression in adult Drosophila melanogaster brain.
1996,
Pubmed
