TY - JOUR
T1 - Mapping of a molecular determinant for protein kinase C β(II) isozyme function
AU - Gökmen-Polar, Yesim
AU - Fields, Alan P.
PY - 1998/8/7
Y1 - 1998/8/7
N2 - In human erythroleukemia (K562) cells, the highly related protein kinase C (PKC) α and PKC β(II) isozymes serve distinct functions in cellular differentiation and proliferation, respectively. Previous studies using two domain switch PKC chimera revealed that the Catalytic domains of PKC α and β(II) contain molecular determinants important for isozyme-specific function (Walker, S. D., Murray, N. R., Burns, D. J., and Fields, A. P. (1995) Proc. Natl. Acad. Sci. U.S.A. 92, 9156-9160). We have now analyzed a panel of PKC chimeras to determine the specific region within the catalytic domain important for PKC β(II) function. A cellular assay for PKC β(II) function was devised based on the finding that PKC β(II) selectively translocates to the nucleus and phosphorylates nuclear lamin B in response to the PKC activator bryostatin. This response is strictly dependent upon expression of PKC β(II) or a PKC chimera that functions like PKC β(II). We demonstrate that a PKC α/β(II) chimera containing only the carboxyl-terminal 13 amino acids from PKC β(II) (β(II) V5) is capable of nuclear translocation and lamin B phosphorylation. These results are consistent with our recent observation that the PKC β(II) V5 region binds to phosphatidylglycerol (PG), a potent and selective PKC β(II) activator present in the nuclear membrane (Murray, N. R., and Fields, A. P. (1998) J. Biol. Chem. 273, 11514-11520). Soluble β(II) V5 peptide selectively inhibits PG-stimulated PKC β(II) activity in a dose-dependent fashion, indicating that PG-mediated activation of PKC β(II) involves interactions with the β(II) V5 region of the enzyme. We conclude that β(II) V5 is a major determinant for PKC β(II) nuclear function and suggest a model in which binding of PG to the β(II) V5 region stimulates nuclear PKC β(II) activity during G2 phase of the cell cycle.
AB - In human erythroleukemia (K562) cells, the highly related protein kinase C (PKC) α and PKC β(II) isozymes serve distinct functions in cellular differentiation and proliferation, respectively. Previous studies using two domain switch PKC chimera revealed that the Catalytic domains of PKC α and β(II) contain molecular determinants important for isozyme-specific function (Walker, S. D., Murray, N. R., Burns, D. J., and Fields, A. P. (1995) Proc. Natl. Acad. Sci. U.S.A. 92, 9156-9160). We have now analyzed a panel of PKC chimeras to determine the specific region within the catalytic domain important for PKC β(II) function. A cellular assay for PKC β(II) function was devised based on the finding that PKC β(II) selectively translocates to the nucleus and phosphorylates nuclear lamin B in response to the PKC activator bryostatin. This response is strictly dependent upon expression of PKC β(II) or a PKC chimera that functions like PKC β(II). We demonstrate that a PKC α/β(II) chimera containing only the carboxyl-terminal 13 amino acids from PKC β(II) (β(II) V5) is capable of nuclear translocation and lamin B phosphorylation. These results are consistent with our recent observation that the PKC β(II) V5 region binds to phosphatidylglycerol (PG), a potent and selective PKC β(II) activator present in the nuclear membrane (Murray, N. R., and Fields, A. P. (1998) J. Biol. Chem. 273, 11514-11520). Soluble β(II) V5 peptide selectively inhibits PG-stimulated PKC β(II) activity in a dose-dependent fashion, indicating that PG-mediated activation of PKC β(II) involves interactions with the β(II) V5 region of the enzyme. We conclude that β(II) V5 is a major determinant for PKC β(II) nuclear function and suggest a model in which binding of PG to the β(II) V5 region stimulates nuclear PKC β(II) activity during G2 phase of the cell cycle.
UR - http://www.scopus.com/inward/record.url?scp=0032493823&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032493823&partnerID=8YFLogxK
U2 - 10.1074/jbc.273.32.20261
DO - 10.1074/jbc.273.32.20261
M3 - Article
C2 - 9685375
AN - SCOPUS:0032493823
SN - 0021-9258
VL - 273
SP - 20261
EP - 20266
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 32
ER -