The cyclic AMP/PKA signal pathway is required for initiation of spore germination in Schizosaccharomyces pombe.
Spore germination, a transition from the quiescent G0 phase to the proliferation cycle, is triggered by glucose in Schizosaccharomyces pombe. The role of cAMP/protein kinase A (PKA) signalling in germination is investigated. Gene disruption of cyr1+, pka1+ and gpa2+ encoding adenylate cyclase, PKA and the alpha-subunit of a trimeric GTP-binding protein, respectively, reduced the colony-forming efficiency of spores in minimal medium. Isolated spores of these null mutants did not germinate in minimal medium for up to 12 h, at which time wild-type spores had completed germination and formed germ projections. In wild-type spores, cortical actin patches randomly distributed in the early stage of outgrowth and then localized to one side of spores before the formation of projections. In contrast, the mutant spores exhibited no actin patches, but the cell surface was predominantly stained, like ungerminated spores of wild-type. Flow fluorocytometric analysis of propidium iodide-stained spores revealed a distinct 1C DNA peak after germination was completed. The fluorescent profile of the mutant spores, however, did not change during 12 h incubation in the minimal medium. These observations indicate that spores harbouring either cyr1Delta, pka1Delta or gpa2Delta are hardly triggered to germination. When wild-type spores were exposed to glucose, the intracellular cAMP level transiently increased in a few minutes, but gpa2Delta spores did not respond to glucose. We conclude that S. pombe spores initiate germination in response to glucose through the cyclic AMP-PKA pathway.
Hatanaka
M
M
Department of Biology, Graduate School of Science, Osaka City University, Sumiyoshi-ku, Osaka 558-8585, Japan.
Shimoda
C
C
D000199:Actins / Q000032:analysis; D000262:Adenylyl Cyclases / Q000235:genetics / Q000502:physiology; D002499:Centrifugation, Density Gradient; D015169:Colony Count, Microbial; D000242:Cyclic AMP / Q000378:metabolism / Q000502:physiology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics / Q000502:physiology*; D004271:DNA, Fungal / Q000032:analysis; D005434:Flow Cytometry; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology; D044385:GTP-Binding Protein alpha Subunits; D015966:Gene Expression Regulation, Fungal; D005947:Glucose / Q000494:pharmacology; D020962:Heterotrimeric GTP-Binding Proteins / Q000235:genetics / Q000502:physiology; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D012568:Schizosaccharomyces / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D029702:Schizosaccharomyces pombe Proteins; D015398:Signal Transduction / Q000235:genetics*; D013172:Spores, Fungal / Q000235:genetics / Q000502:physiology
D000199:Actins; D004271:DNA, Fungal; D005656:Fungal Proteins; D044385:GTP-Binding Protein alpha Subunits; D029702:Schizosaccharomyces pombe Proteins; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D020962:Heterotrimeric GTP-Binding Proteins; C540695:gpa2 protein, S pombe; D000262:Adenylyl Cyclases; D005947:Glucose