CG15031/PPYR1 is an intrinsically unstructured protein that interacts with protein phosphatase Y.
Protein phosphatase Y (PPY) is a Drosophila testis-specific enzyme of unknown function. In a yeast two-hybrid screen we identified CG15031/PPYR1 as a PPY interacting protein. The specificity of the protein-protein interaction was proven by directed two-hybrid tests. The complex formation between PPY and PPYR1 was confirmed under in vitro and in vivo conditions by plasmon resonance spectroscopy, co-immunoprecipitation, and pull down experiments. Recombinant PPYR1 expressed in Escherichia coli is a heatstable, protease sensitive, intrinsically unstructured RNA-binding protein that migrates anomalously in SDS-polyacrylamide gel electrophoresis. It can be phosphorylated by cAMP-dependent protein kinase in vitro. PPYR1 moderately inhibits PPY activity, the inhibitory potential of the protein is slightly increased by phosphorylation. We suggest that PPYR1 may function as a scaffolding protein that targets PPY to RNA and other protein partners in Drosophila melanogaster.
Kókai
Endre
E
Department of Medical Chemistry, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, H-4032 Debrecen, Egyetem tér 1, Hungary.
Tantos
Agnes
A
Vissi
Emese
E
Szöor
Balázs
B
Tompa
Péter
P
Gausz
János
J
Alphey
Luke
L
Friedrich
Péter
P
Dombrádi
Viktor
V
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D016601:RNA-Binding Proteins / Q000378:metabolism; D017476:Receptors, Neuropeptide Y / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020349:Surface Plasmon Resonance
D029721:Drosophila Proteins; C512449:PPYR1 protein, Drosophila; D016601:RNA-Binding Proteins; D017476:Receptors, Neuropeptide Y; D011994:Recombinant Proteins; C109491:neuropeptide Y4 receptor; D017868:Cyclic AMP-Dependent Protein Kinases; C104054:PpY-55A protein, Drosophila; D010749:Phosphoprotein Phosphatases