Ras-dependent signal transduction is indispensable but not sufficient for the activation of AP1/Jun by PKC delta.
Modulation of gene expression by 12-O-tetradecanoylphorbol-13-acetate (TPA) is thought to be mediated by protein kinase C (PKC), a major cellular receptor for TPA. We confirm this by showing that the overexpression of PKC delta enhances the TPA induction of the TRE-tk-CAT reporter gene in NIH3T3 cells. To investigate the mutual relationship between PKC delta- and Ras-dependent signal transduction pathways to a TRE binding transcription factor, AP1/Jun, we constructed constitutively active and dominant negative mutants of PKC delta. Activated Ras induced reporter gene expression in collaboration with overexpressed c-Jun or JunD, and this induction was insensitive to the dominant negative PKC delta. On the other hand, reporter gene expression induced by the constitutively active PKC delta was severely inhibited by dominant negative Ras, as well as by the dominant negative PKC delta. Thus, Ras activation must be indispensable for PKC delta to activate AP1/Jun. In the absence of overexpressed c-Jun or JunD, activated Ras was, however, clearly less effective than constitutively active PKC delta which showed full activation of reporter gene expression by itself. This suggests the presence of an additional, Ras-independent, signaling pathway downstream of PKC delta to activate AP1/Jun. In spite of the remarkable ability of constitutively active PKC delta to activate TRE-tk-CAT expression, this mutant suppressed cell growth.
Hirai
S
S
Department of Molecular Biology, Yokohama City University, School of Medicine, Japan.
Izumi
Y
Y
Higa
K
K
Kaibuchi
K
K
Mizuno
K
K
Osada
S
S
Suzuki
K
K
Ohno
S
S
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002455:Cell Division; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D011905:Genes, ras; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008957:Models, Genetic; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D051745:Protein Kinase C-delta; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D015398:Signal Transduction
D007527:Isoenzymes; D016755:Proto-Oncogene Proteins c-jun; D011993:Recombinant Fusion Proteins; D015500:Chloramphenicol O-Acetyltransferase; C497259:Prkcd protein, mouse; D011493:Protein Kinase C; D051745:Protein Kinase C-delta