The SH3 domain of the S. cerevisiae Cdc25p binds adenylyl cyclase and facilitates Ras regulation of cAMP signalling.
Cdc25 and Ras are two proteins required for cAMP signalling in the budding yeast Saccharomyces cerevisiae. Cdc25 is the guanine nucleotide exchange protein that activates Ras. Ras, in turn, activates adenylyl cyclase. Cdc25 has a Src homology 3 (SH3) domain near the N-terminus and a catalytic domain in the C-terminal region. We find that a point mutation in the SH3 domain attenuates cAMP signalling in response to glucose feeding. Furthermore, we demonstrate, by using recombinant adenylyl cyclase and Cdc25, that the SH3 domain of Cdc25 can bind directly to adenylyl cyclase. Binding was specific, because the SH3 domain of Abp1p (actin-binding protein 1), which binds the 70,000 Mr subunit of adenylyl cyclase, CAP/Srv2, failed to bind adenylyl cyclase. A binding site for Cdc25-SH3 localised to the C-terminal catalytic region of adenylyl cyclase. Finally, pre-incubation with Ras enhanced the SH3-bound adenylyl cyclase activity. These studies suggest that a direct interaction between Cdc25 and adenylyl cyclase promotes efficient assembly of the adenylyl cyclase complex.
Mintzer
K A
KA
Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia 19104, USA.
Field
J
J
D000262:Adenylyl Cyclases / Q000378:metabolism*; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D000242:Cyclic AMP / Q000032:analysis / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D015870:Gene Expression; D005947:Glucose / Q000494:pharmacology; D008274:Magnesium / Q000378:metabolism; D008954:Models, Biological; D016296:Mutagenesis; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000378:metabolism*; D017354:Point Mutation; D011485:Protein Binding; D011993:Recombinant Fusion Proteins; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D015398:Signal Transduction; D013997:Time Factors; D015003:Yeasts; D018631:ras Proteins / Q000502:physiology*; D020666:ras-GRF1; D018909:src Homology Domains / Q000502:physiology*
D018797:Cell Cycle Proteins; D011993:Recombinant Fusion Proteins; D020666:ras-GRF1; D000242:Cyclic AMP; D010749:Phosphoprotein Phosphatases; D018631:ras Proteins; D000262:Adenylyl Cyclases; D008274:Magnesium; D005947:Glucose