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doi:10.1534/genetics.105.047233
A more recent version of this article appeared on December 1, 2005.
REGULAR RESEARCH PAPERS |
Schizosaccharomyces pombe adenylate cyclase suppressor mutations suggest a role for cAMP phosphodiesterase regulation in feedback control of glucose/cAMP signaling
Lili Wang 1, Kenneth Griffiths 1, Y. Hi Zhang 1, F. Douglas Ivey 1 and Charles S. Hoffman 1*
1 Boston College
* To whom correspondence should be addressed. E-mail: hoffmacs{at}bc.edu.
Submitted on June 23, 2005
Revised on July 29, 2005
Accepted on 18 August 2005
Mutations affecting the Schizosaccharomyces pombe cAMP phosphodiesterase gene cgs2+ were identified in a screen for suppressors of mutant alleles of the adenylate cyclase gene (git2+/cyr1+), which encode catalytically active forms of the enzyme that cannot be stimulated by extracellular glucose signaling. These mutations suppress both of the git2- mutant alleles used in the suppressor selection, as well as mutations in git1+, git3+, git5+, git7+, git10+, and git11+, which are all required for adenylate cyclase activation. Notably, these cgs2 mutant alleles fail to suppress mutations in gpa2+, which encodes the G? subunit of a heterotrimeric G protein required for adenylate cyclase activation, although the previously identified cgs2-2 allele does suppress loss of gpa2+. Further analysis of the cgs2-s1 allele reveals a synthetic interaction with the gpa2R176H activated allele, with respect to derepression of fbp1-lacZ transcription in glucose-starved cells. In addition, direct measurements of cAMP levels show that cgs2-s1 cells maintain normal basal cAMP levels, but are severely defective in feedback regulation upon glucose detection. These results suggest that PDE activity in S. pombe may be coordinately regulated with adenylate cyclase activity as part of the feedback regulation mechanism to limit the cAMP response to glucose detection.
Key Words: Schizosaccharomyces pombe, adenylate cyclase, cAMP phosphodiesterase, cgs2, git2
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