Non-histone chromatin proteins in beef thyroid: distinct phosphorylation patterns of several protein kinases.
Non-histone chromatin protein (NHCP) fractions were extracted from purified beef thyroid nuclear preparations and tested for the presence of protein kinase activities using several known mediators of thyroid regulation, as well as potential phosphotransferase substrates using purified or partially purified protein kinase activities. The addition of cAMP/3-isobutyl-1-methylxanthine had no effect on NHCP histone kinase activity; the addition of 10 micrograms of the heat-stable cAMP-dependent protein kinase A inhibitor, however, resulted in a 47% reduction in histone H1 kinase activity. Nuclear casein kinase II activity was present in the NHCP fractions as evidenced by the capacity of spermine to stimulate (ED50 = 0.19 mM) and heparin to inhibit (ID50 = 0.09 microgram/ml) the phosphorylation of casein; further, the phosphotransferase activity could be purified by sequential casein-agarose and spermine-agarose affinity chromatography. Neither calcium-calmodulin nor calcium/phosphatidylserine/diolein had an effect on NHCP casein kinase or histone kinase activities, respectively. The addition of cAMP-dependent protein kinase A catalytic subunit, nuclear casein kinase II, calcium-activated calmodulin-dependent protein kinase and diacylglycerol-activated calcium/phospholipid-dependent protein kinase C activities exhibited distinct phosphorylation patterns when NHCP were used as substrates and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. We conclude that NHCP fraction from beef thyroid: 1) contains both cAMP-dependent protein kinase A catalytic subunit and nuclear casein kinase II and 2) substrates for cAMP-dependent protein kinase A, calcium-activated calmodulin-dependent protein kinase, protein kinase C, and nuclear casein kinase II.
Levasseur
S
S
Department of Medicine, Cook County Hospital, Chicago, Illinois 60612.
Poleck
T
T
Friedman
Y
Y
Burke
G
G
D000818:Animals; D047388:Casein Kinases; D002417:Cattle; D002843:Chromatin / Q000378:metabolism*; D010766:Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism; D006656:Protamine Kinase / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism*; D013961:Thyroid Gland / Q000378:metabolism*
D002843:Chromatin; D010770:Phosphotransferases; D011494:Protein Kinases; D047388:Casein Kinases; D006656:Protamine Kinase