Dictyostelium cells express an aggregative adenylylcyclase (ACA), resp
onsible for oscillatory cAMP signaling, and a spore germination-specif
ic adenylycyclase (ACG), Overexpression of PKA regulatory (R) subunits
blocks oscillatory cAMP signaling but increases basal cAMP levels, wh
ile neither ACA nor ACG mRNA could be detected, To test whether a nove
l type of adenylylcyclase (AC) was responsible for cAMP synthesis, dom
inant-negative PKA-R subunits (PKA-R(M)) and control R-subunits (PKA-R
(c)) were overexpressed in ACA null mutants, Both transformations as w
ell as transformation with an unrelated vector, carrying the same NEO(
R) selection marker, induced an AC activity in growing cells with the
biochemical characteristics of ACG. Similar vectors with a different U
RA selection marker did not increase AC activity. We conclude that the
aminoglycoside phosphotransferase encoded by the very commonly used N
EO(R) selection marker induces ectopic ACG; activity in Dictyostelium
cells, (C) 1995 Academic Press, Inc.