Protein phosphatase 2A reverses phosphorylation of c-Jun specified by the delta domain in vitro: correlation with oncogenic activation and deregulated transactivation activity of v-Jun.
Chicken c-Jun proteins synthesized in vitro in reticulocyte extract consist of several electrophoretic isoforms resulting from phosphorylation which can be specifically reversed by purified protein phosphatase 2A (PP2A). Using the phosphatase inhibitors okadaic acid and microcystin-LR, we conclude that the isoforms seen in vitro represent a balance between the action of an unidentified kinase(s) which phosphorylates c-Jun and dephosphorylation by an endogenous PP2A-like phosphatase. c-Jun proteins are also subject to phosphorylation in vivo in chick embryo fibroblasts (CEF), which can be reversed by PP2A. In contrast, the viral Jun oncoprotein encoded by ASV17 is not subject to PP2A-sensitive phosphorylation in vitro and is hypophosphorylated in comparison with c-Jun in ASV17-transformed CEF. Hybrids between c-Jun and v-Jun demonstrate that differential phosphorylation in vitro is a consequence of deletion of 27 amino acids in the N-terminal third of v-Jun. The deletion is important for oncogenic activation and lies in a domain, termed delta, which regulates c-Jun transactivation function. PP2A-sensitive phosphorylation in vitro correlates with the differential responsiveness of c-Jun and v-Jun to a recently identified cell type-specific inhibitor of transactivation function.
Black
E J
EJ
Cancer Research Campaign Beatson Laboratories, Beatson Institute for Cancer Research, Bearsden, Glasgow, UK.
Street
A J
AJ
Gillespie
D A
DA
D000595:Amino Acid Sequence; D000818:Animals; D002645:Chickens; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004988:Ethers, Cyclic / Q000494:pharmacology; D016758:Genes, jun; D007132:Immunoglobulin Isotypes / Q000737:chemistry; D008387:Marine Toxins; D051379:Mice; D052998:Microcystins; D008969:Molecular Sequence Data; D019319:Okadaic Acid; D010456:Peptides, Cyclic / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000494:pharmacology* / Q000502:physiology; D010766:Phosphorylation / Q000187:drug effects; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D054648:Protein Phosphatase 2; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D016755:Proto-Oncogene Proteins c-jun / Q000032:analysis; D012323:RNA Processing, Post-Transcriptional; D014158:Transcription, Genetic
D004988:Ethers, Cyclic; D007132:Immunoglobulin Isotypes; D008387:Marine Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2; C057862:cyanoginosin LR