Calmodulin can modulate protein phosphorylation in rat liver cells nuclei.
This report describes the immunological identification of a 60-kDa calmodulin-binding protein, previously detected in the nuclei of rat liver cells (Bachs, O., Lanini, L., Serratosa, J., Coll, M.J., Bastos, R., Aligué, R., Rius, E., and Carafoli, E. (1990) J. Biol. Chem. 265, 18595-18600), as the calmodulin-dependent protein phosphatase calcineurin. Calcineurin could be extracted from the nuclei by incubation with DNase and RNase, indicating that it is associated with nuclear structures sensitive to the action of nucleases (chromatin or/and ribonucleoproteins). The presence of calcineurin in the nuclei of rat liver cells indicates that calmodulin may modulate the phosphorylation level of nuclear proteins by promoting their dephosphorylation. This report also shows that calmodulin inhibits the activity of casein kinase-2 in the nuclear fractions obtained by nuclease extraction. Phosphorylation experiments indicate that casein kinase-2 phosphorylates three major substrates of 100, 42-44, and 37 kDa as well as other minor proteins in the nuclease extracts. Calmodulin reduces the phosphorylation level of the two latter major proteins and of a minor band of 50 kDa. Thus, nuclear calmodulin in rat liver cells could regulate phosphorylation of nuclear proteins by at least two mechanisms: 1) activation of calcineurin and 2) inhibition of casein kinase-2.
Department of Cell Biology, Faculty of Medicine, University of Barcelona, Spain.
R
R
Bosser
R
R
Aligué
D
D
Guerini
N
N
Agell
E
E
Carafoli
O
O
Bachs
D000818:Animals; D015153:Blotting, Western; D019703:Calcineurin; D002147:Calmodulin / Q000378:metabolism*; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D047388:Casein Kinases; D002417:Cattle; D002467:Cell Nucleus / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D007457:Iodine Radioisotopes; D008099:Liver / Q000378:metabolism*; D008297:Male; D009687:Nuclear Proteins / Q000378:metabolism*; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D007457:Iodine Radioisotopes; D009687:Nuclear Proteins; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases; D047388:Casein Kinases; D019703:Calcineurin; D010749:Phosphoprotein Phosphatases