Journal of Neuroscience, Vol 12, 1628-1639, Copyright © 1992 by Society for Neuroscience
Analysis of cis-regulatory elements in the 5' flanking region of the Drosophila melanogaster choline acetyltransferase gene
T Kitamoto, K Ikeda and PM Salvaterra
Division of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, California 91010.
We have analyzed the cis-regulatory elements in the 5' flanking region of
the Drosophila choline acetyltransferase gene (ChAT, E.C.2.3.1.6). DNA
fragments were fused to the Escherichia coli lacZ reporter gene and
introduced into the Drosophila germ line by P-element-mediated
transformation. A 7.4 kb 5' flanking sequence directed beta- galactosidase
expression in the adult optic lobes and other well- defined CNS structures
with a pattern very similar to the distribution of endogenous ChAT protein.
In contrast, the proximal 3.3 kb and 1.2 kb of 5' flanking DNA directed
lacZ expression in only selected subsets of the structures seen with the
7.4 kb lacZ construct. Our results indicate that both qualitative and
quantitative regulatory elements are present in the 5' flanking DNA and
that these elements distinguish various subsets of cholinergic neurons. We
have also fused the same 5' flanking DNA sequences to wild-type ChAT cDNA
and used these constructs to transform Chatsl mutant flies. Not only the
7.4 kb cDNA construct, but also the 3.3 and 1.2 kb constructs, rescued
Chatsl from temperature- dependent paralysis and adult lethality,
indicating that the regulatory information in any of these genomic
fragments can drive sufficient wild- type ChAT expression to overcome these
mutant phenotypes.
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