Deletions in the regulatory or kinase domains of protein kinase C-alpha cause association with the cell nucleus.
We have constructed expression plasmids carrying protein kinase C (PKC) cDNAs with deletions in the coding region. Two truncated molecules, consisting only of the kinase domain of PKC-alpha, were generated by removing parts of the cDNA coding for the regulatory region. Another mutant molecule was created by deleting 95 amino acids from the C-terminal part of the molecule. The full-length cDNA coding for PKC-alpha and its deletion constructs was expressed in COS cells. Using cell fractionation experiments and immunofluorescence staining, we demonstrate here that in contrast to the cytosolic localization of full-length PKC-alpha, the truncated forms, coding only for the kinase domain, were found exclusively in the cell nucleus. Further subfractionation of nuclei isolated from these transfected cells indicated partial association with the nuclear envelopes. Expression of the cDNA lacking the C-terminal part of the molecule in COS cells encoded a truncated molecule that was found both in the cytosol and in the nucleus. We also show that translocation of full-length PKC-alpha molecules to the cell nucleus occurred in response to phorbol ester treatment. Thus, it appears that accumulation of PKC-alpha in the nucleus results either by phorbol ester activation or by deletions of specific regions of the molecule. A molecular mechanism for the nuclear translocation of phorbol ester-activated PKC-alpha or its truncated molecules is suggested.
Eldar
H
H
Department of Chemical Immunology, Weizmann Institute of Science, Rehovot, Israel.
Ben-Chaim
J
J
Livneh
E
E
D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport; D015153:Blotting, Western; D002460:Cell Line; D002467:Cell Nucleus / Q000201:enzymology*; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D008969:Molecular Sequence Data; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection
D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate