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Volume 17, Number 22, Issue of November 15, 1997 pp. 8817-8827

Defective Learning in Mutants of the Drosophila Gene for a Regulatory Subunit of cAMP-Dependent Protein Kinase

Received April 9, 1997; revised Aug. 27, 1997; accepted Aug. 29, 1997.

Stephen F. Goodwin1, Maria Del Vecchio2, Klara Velinzon2, Catherine Hogel2, Steven R. H. Russell3, Tim Tully2, and Kim Kaiser1

1 Institute of Genetics, University of Glasgow, Glasgow G11 5JS, Scotland, 2 Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, and 3 Department of Genetics, University of Cambridge, Cambridge CB2 3EH, England

Disruptions of a Drosophila gene encoding a regulatory subunit of cAMP-dependent protein kinase homologous to mammalian RIbeta (dPKA-RI) were targeted to the first (noncoding) exon of dPKA-RI via site-selected P element mutagenesis. Flies homozygous for either of two mutant alleles showed specific defects in olfactory learning but not in subsequent memory decay. In contrast, olfactory acuity and shock reactivity, component behaviors required for normal odor avoidance learning, were normal in these mutants. Northern and Western blot analyses of mRNA and protein extracted from adult heads have revealed a complex lesion of the PKA-RI locus, including expression of a novel product and over- or underexpression of wild-type products in mutants. Western blot analysis revealed reductions in RI protein in mutants. PKA activity in the absence of exogenous cAMP also was significantly higher than normal in homogenates from mutant adult heads. These two mutant alleles failed to complement each other for each of these phenotypic defects, eliminating second-site mutations as a possible explanation. These results establish a role for an RI regulatory subunit of PKA in Pavlovian olfactory conditioning.

Key words: learning; memory; reverse genetics; PKA; insertional mutagenesis; RI regulatory subunit




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