Casein kinase 2 and protein substrates are released from rat liver cells nuclei by DNase or RNase digestion.
Casein kinase 2 was released from rat liver cells nuclei by digestion with DNase I plus RNase A. This treatment also released three major substrates of 50, 40-42, and 37 kDa. Casein kinase 2 and substrates were also extracted by DNase or RNase separately. However, in DNase extracts only the 37 kDa protein was phosphorylated by casein kinase 2, whereas in RNase extracts all three substrates were phosphorylated. When the DNase extracts were subsequently treated with RNase the 40-42 substrates were then phosphorylated, indicating that their interaction with RNA prevents their phosphorylation by casein kinase 2. The ratio of B: alpha subunits of casein kinase 2 present in the nuclease extracts was higher than that of the purified enzyme, which is assumed to be 1:1. A further analysis by sucrose gradient centrifugation revealed that under physiological salt conditions casein kinase 2 from nuclease extracts formed large aggregates (higher than 300 kDa) which were disrupted at 400 mM KCl. At the latter KCl concentration CK-2 activity was localized at a position corresponding to a M(r) of 230-250 kDa, which is still higher than the typical tetrameric form of the enzyme.
Bosser
R
R
Department of Cell Biology, Faculty of Medicine, University of Barcelona, Spain.
Roig
J
J
Itare
E
E
Bachs
O
O
D000818:Animals; D047388:Casein Kinases; D002467:Cell Nucleus / Q000201:enzymology*; D002499:Centrifugation, Density Gradient; D003850:Deoxyribonuclease I / Q000378:metabolism*; D008099:Liver / Q000648:ultrastructure*; D008970:Molecular Weight; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011189:Potassium Chloride; D011494:Protein Kinases / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D012259:Ribonuclease, Pancreatic / Q000378:metabolism*
D010750:Phosphoproteins; D011506:Proteins; D011189:Potassium Chloride; D011494:Protein Kinases; D047388:Casein Kinases; D003850:Deoxyribonuclease I; D012259:Ribonuclease, Pancreatic