Cellular compartmentalization of protein kinase activity during the cell cycle.
The quantities and types of protein kinases found in the cytoplasmic and nuclear or chromosomal compartments of interphase and mitotic human culture cells were compared. Using histone as substrate, the total quantity of kinases recovered from cytoplasmic and chromosomal fractions of mitotic cells was several times greater than from cytoplasmic and nuclear fractions of interphase cells. In both mitotic and interphase cells, more activity was recovered from cytoplasmic fractions than from chromosomal or nuclear fractions, respectively. When activity against various substrates was examined, mitotic chromosomal extracts were found to display the greatest preference for the H1 fraction of histones. Neither cytoplasmic nor chromosomal fractions from mitotic cells exhibited enhanced activity in the presence of cAMP, whereas the activity of both cytoplasmic and nuclear fractions of interphase cells was enhanced. Protein kinases, previously identified by nondenaturing polyacrylamide gel electrophoresis as present in the cytoplasmic fraction of mitotic but not interphase cells, were also present in chromosomal fractions of mitotic cells; only one of these kinases may be present in nuclear extracts of interphase cells. In addition, the profiles of nuclear extracts of interphase cells differ from their cytoplasmic fractions. These results indicate that there are protein kinases which are restricted to the mitotic phase of the cell cycle and that they apparently partition between the cytoplasmic and chromosomal compartments of cells in mitosis.
Fabisz-Kijowska
A A
AA
Department of Molecular and Cell Biology, Pennsylvania State University, University Park 16802.
Lumley-Sapanski
K
K
Halleck
M S
MS
Schlegel
R A
RA
D002451:Cell Compartmentation; D002453:Cell Cycle; D002457:Cell Extracts; D002461:Cell Line, Transformed; D005591:Chemical Fractionation; D000242:Cyclic AMP / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D011506:Proteins / Q000032:analysis; D013379:Substrate Specificity
D002457:Cell Extracts; D011506:Proteins; D000242:Cyclic AMP; D011494:Protein Kinases