Suppression of mitogenic activity by stable expression of the regulatory domain of PKC beta.
The amino-terminal regulatory domain portion of each protein kinase C (PKC) family member (which in the case of PKC beta 1 includes the pseudosubstrate, C1, V1 and C2 domains) plays an important role in regulating the kinase activity of the carboxyl-terminal catalytic domain. To examine the possibility that this regulatory domain region (designated 'PAT') might have biological functions independent of the catalytic domain, we have developed derivatives of R6 cells which stably express a truncated PKC beta 1 cDNA that encodes the amino-terminal 317 amino acids, including the entire regulatory domain. These R6-plPAT cells express abundant amounts of a 38 kDa protein which binds a labeled phorbol ester, but lacks protein kinase activity. In contrast to the 79 kDa PKC beta 1 holoenzyme which, when overexpressed in R6 cells, is found mostly in the cytosol, the 38 kDa PAT protein is predominantly associated with the particulate subcellular fraction. Furthermore, the PAT protein fails to show down-regulation following treatment of R6-plPAT cells with 12-O-tetradecanoylphorbol-13-acetate (TPA). Evidence is also presented that TPA-stimulated growth is suppressed in R6-plPAT cells. These findings suggest that the PKC beta 1 regulatory domain could be involved in the suppression of mitogenic signaling.
Kahn
S M
SM
Columbia Presbyterian Cancer Center, Columbia University, New York, NY 10032.
O'Driscoll
K R
KR
Jiang
W
W
Borner
C
C
Xu
D B
DB
Blackwood
M A
MA
Zhang
Y J
YJ
Nomoto
K
K
Weinstein
I B
IB
D000818:Animals; D001483:Base Sequence; D002455:Cell Division; D002460:Cell Line; D003001:Cloning, Molecular; D003260:Contact Inhibition; D003600:Cytosol / Q000201:enzymology; D004790:Enzyme Induction / Q000187:drug effects; D005347:Fibroblasts; D007527:Isoenzymes / Q000096:biosynthesis / Q000737:chemistry*; D008938:Mitosis / Q000502:physiology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000096:biosynthesis*; D011493:Protein Kinase C / Q000096:biosynthesis / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D015398:Signal Transduction / Q000502:physiology*; D013347:Subcellular Fractions / Q000201:enzymology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D007527:Isoenzymes; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate