title: Regulated protein degradation controls PKA function and cell-type differentiation in Dictyostelium.
abstract: Cullins function as scaffolds that, along with F-box/WD40-repeat-containing proteins, mediate the ubiquitination of proteins to target them for degradation by the proteasome. We have identified a cullin CulA that is required at several stages during Dictyostelium development. culA null cells are defective in inducing cell-type-specific gene expression and exhibit defects during aggregation, including reduced chemotaxis. PKA is an important regulator of Dictyostelium development. The levels of intracellular cAMP and PKA activity are controlled by the rate of synthesis of cAMP and its degradation by the cAMP-specific phosphodiesterase RegA. We show that overexpression of the PKA catalytic subunit (PKAcat) rescues many of the culA null defects and those of cells lacking FbxA/ChtA, a previously described F-box/WD40-repeat-containing protein, suggesting CulA and FbxA proteins are involved in regulating PKA function. Whereas RegA protein levels drop as the multicellular organism forms in the wild-type strain, they remain high in culA null and fbxA null cells. Although PKA can suppress the culA and fbxA null developmental phenotypes, it does not suppress the altered RegA degradation, suggesting that PKA lies downstream of RegA, CulA, and FbxA. Finally, we show that CulA, FbxA, and RegA are found in a complex in vivo, and formation of this complex is dependent on the MAP kinase ERK2, which is also required for PKA function. We propose that CulA and FbxA regulate multicellular development by targeting RegA for degradation via a pathway that requires ERK2 function, leading to an increase in cAMP and PKA activity.
authors.affiliation: Section of Cell and Developmental Biology and Center for Molecular Genetics, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: S
authors.initials: S
authors.lastname: Mohanty
authors.forename: S
authors.initials: S
authors.lastname: Lee
authors.forename: N
authors.initials: N
authors.lastname: Yadava
authors.forename: M J
authors.initials: MJ
authors.lastname: Dealy
authors.forename: R S
authors.initials: RS
authors.lastname: Johnson
authors.forename: R A
authors.initials: RA
authors.lastname: Firtel
mesh_terms: D015105:3',5'-Cyclic-AMP Phosphodiesterases; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism*; D004023:Dictyostelium / Q000166:cytology* / Q000201:enzymology* / Q000235:genetics; D044783:F-Box Proteins; D018507:Gene Expression Regulation, Developmental; D019950:Mitogen-Activated Protein Kinase 1 / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009024:Morphogenesis; D009154:Mutation; D016366:Open Reading Frames; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D014452:Ubiquitins / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D018797:Cell Cycle Proteins; C406731:ChtA protein, Dictyostelium; C429923:CulA protein, Dictyostelium; D044783:F-Box Proteins; C412452:FbxA protein, Dictyostelium; D015800:Protozoan Proteins; D014452:Ubiquitins; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D019950:Mitogen-Activated Protein Kinase 1; D015105:3',5'-Cyclic-AMP Phosphodiesterases; C112742:regA protein, Dictyostelium