Importance of C1B Domain for Lipid Messenger-induced Targeting of Protein Kinase C

Abstract

The molecular mechanisms by which arachidonic acid (AA) and ceramide elicit translocation of protein kinase C (PKC) were investigated. Ceramide translocated εPKC from the cytoplasm to the Golgi complex, but with a mechanism distinct from that utilized by AA. Using fluorescence recovery after photobleaching, we showed that, upon treatment with AA, εPKC was tightly associated with the Golgi complex; ceramide elicited an accumulation of εPKC which was exchangeable with the cytoplasm. Stimulation with ceramide after AA converted the AA-induced Golgi complex staining to one elicited by ceramide alone; AA had no effect on the ceramide-stimulated localization. Using point mutants and deletions of εPKC, we determined that the εC1B domain was responsible for the ceramide- and AA-induced translocation. Switch chimeras, containing the C1B from εPKC in the context of δPKC (δ(εC1B)) and vice versa (ε(δC1B)), were generated and tested for their translocation in response to ceramide and AA. δ(εC1B) translocated upon treatment with both ceramide and AA; ε(δC1B) responded only to ceramide. Thus, through the C1B domain, AA and ceramide induce different patterns of εPKC translocation and the C1B domain defines the subtype specific sensitivity of PKCs to lipid second messengers. The molecular mechanisms by which arachidonic acid (AA) and ceramide elicit translocation of protein kinase C (PKC) were investigated. Ceramide translocated εPKC from the cytoplasm to the Golgi complex, but with a mechanism distinct from that utilized by AA. Using fluorescence recovery after photobleaching, we showed that, upon treatment with AA, εPKC was tightly associated with the Golgi complex; ceramide elicited an accumulation of εPKC which was exchangeable with the cytoplasm. Stimulation with ceramide after AA converted the AA-induced Golgi complex staining to one elicited by ceramide alone; AA had no effect on the ceramide-stimulated localization. Using point mutants and deletions of εPKC, we determined that the εC1B domain was responsible for the ceramide- and AA-induced translocation. Switch chimeras, containing the C1B from εPKC in the context of δPKC (δ(εC1B)) and vice versa (ε(δC1B)), were generated and tested for their translocation in response to ceramide and AA. δ(εC1B) translocated upon treatment with both ceramide and AA; ε(δC1B) responded only to ceramide. Thus, through the C1B domain, AA and ceramide induce different patterns of εPKC translocation and the C1B domain defines the subtype specific sensitivity of PKCs to lipid second messengers. The PKC 1The abbreviations used are: PKCprotein kinase CAAarachidonic acidFRAPfluorescent recovery after photobleachingGFPgreen fluorescent proteinεPKCε subtype of protein kinase CδPKCδ subtype of protein kinase CTPA12-O-tetradecanoylphorbol-13-acetateWGAwheat germ agglutininDGdiacylglycerolPSpseudosubstrateC1conserved region 1GFPgreen fluorescent proteinCHOChinese hamster ovaryPBSphosphate-buffered salinePDBuphorbol 12,13-dibutyrate 1The abbreviations used are: PKCprotein kinase CAAarachidonic acidFRAPfluorescent recovery after photobleachingGFPgreen fluorescent proteinεPKCε subtype of protein kinase CδPKCδ subtype of protein kinase CTPA12-O-tetradecanoylphorbol-13-acetateWGAwheat germ agglutininDGdiacylglycerolPSpseudosubstrateC1conserved region 1GFPgreen fluorescent proteinCHOChinese hamster ovaryPBSphosphate-buffered salinePDBuphorbol 12,13-dibutyrate family of serine/threonine protein kinase contains at least 10 subtypes. They are divided into three subgroups based on structural differences and requirement for activators (1Nishizuka Y. Nature. 1984; 308: 693-698Crossref PubMed Scopus (5740) Google Scholar, 2Nishizuka Y. Nature. 1988; 334: 661-665Crossref PubMed Scopus (3523) Google Scholar, 3Nishizuka Y. Science. 1992; 258: 607-614Crossref PubMed Scopus (4214) Google Scholar, 4Nishizuka Y. FASEB J. 1995; 9: 484-496Crossref PubMed Scopus (2352) Google Scholar). The conventional PKCs (cPKC; α, βI, βII, and γ) are Ca2+-dependent and activated by diacylglycerol or phorbol esters. The novel PKCs (nPKC; δ, ε, η, and θ) are activated by diacylglycerol (DG) or phorbol esters, but are Ca2+-independent (5Ono Y. Fujii T. Igarashi K. Kikkawa U. Ogita K. Nishizuka Y. Nucleic Acids Res. 1988; 16: 5199-5200Crossref PubMed Scopus (58) Google Scholar, 6Osada S. Mizuno K. Saido T.C. Suzuki K. Kuroki T. Ohno S. Mol. Cell. Biol. 1992; 12: 3930-3938Crossref PubMed Google Scholar, 7Hug J. PubMed Scopus Google Scholar). The PKCs and are to and are Ca2+-independent Y. Fujii T. Igarashi K. T. Kikkawa U. Nishizuka Y. U. S. PubMed Scopus Google Scholar, K. Mizuno K. S. S. S. Suzuki K. Ohno S. J. Biol. PubMed Google Scholar, Y. J. Biol. PubMed Google PKCs an domain and a domain in the The domain of the PKCs contains a region a and a region The of εPKC to a of εPKC translocation J. Biol. PubMed Scopus Google Scholar, T. J. J. Biol. PubMed Scopus Google Scholar). the the in the of the domain, the by the The of activators to the domain a which the from the and the Biol. 9: PubMed Scopus Google Scholar). and phorbol to the domain Y. Nature. 1988; 334: 661-665Crossref PubMed Scopus (3523) Google Scholar, Y. Fujii T. Igarashi K. T. Kikkawa U. Nishizuka Y. U. S. PubMed Scopus Google Scholar, Biol. 1995; PubMed Scopus Google Scholar, J. Biol. PubMed Google Scholar). the domain that of εPKC J. Biol. PubMed Scopus Google Scholar, S. Mol. Biol. Cell. PubMed Scopus Google Scholar, Suzuki K. J. Y. T. J. Biol. PubMed Scopus Google domain of and and of and in a The C1B of and showed phorbol but for showed for phorbol K. K. Y. Kikkawa U. J. Scopus Google Scholar). or translocated to the in response to or phorbol translocation of C1B was only in phorbol T. J. Biol. PubMed Scopus Google Scholar). distinct for the and C1B in the of the J. Biol. PubMed Scopus Google Scholar). that the and C1B of PKCs are of PKC only by and but by arachidonic acid (AA) K. Kikkawa U. J. 1995; PubMed Scopus Google Scholar, T. Y. K. Nishizuka Y. U. S. PubMed Scopus Google and ceramide J. PubMed Scopus Google Scholar, J. Res. PubMed Scopus Google Scholar). esters, lipid second induce translocation of and in that AA translocation of εPKC Y. J. Google and ceramide εPKC and δPKC from the to the cytoplasm T. Y. S. J. Biol. PubMed Scopus Google Scholar). Using fluorescent protein PKCs and we that AA εPKC, but from the cytoplasm to the Golgi complex Y. K. K. J. Biol. PubMed Scopus Google Scholar, Y. K. J. Google and that ceramide δPKC from the cytoplasm to the Golgi complex T. S. Y. Mol. Cell. Biol. PubMed Scopus Google Scholar). the mechanism we the of and δPKC that to ceramide and AA to the molecular mechanisms responsible for the translocation of of ceramide and AA lipid only to Ceramide in T. J. Biol. PubMed Google of Y. S. J. PubMed Google T. Y. S. J. Biol. PubMed Scopus Google Scholar, Science. PubMed Scopus Google Scholar, Science. PubMed Scopus Google and of PubMed Scopus Google Scholar). AA to a J. Nature. 1992; PubMed Scopus Google in the of a of T. Res. PubMed Scopus Google a of J. Res. PubMed Scopus Google Scholar, J. Biol. PubMed Scopus Google and to for J. Biol. PubMed Scopus Google Scholar). ceramide and AA the translocation of AA εPKC but δPKC in K. Kikkawa U. J. 1995; PubMed Scopus Google and the of J. Res. PubMed Scopus Google Scholar). S. J. Biol. PubMed Scopus Google that and δPKC translocated by of AA. T. J. Biol. PubMed Scopus Google that AA translocation of the domain of Ceramide δPKC and εPKC from the to the cytoplasm T. Y. S. J. Biol. PubMed Scopus Google and from the cytoplasm to the J. PubMed Scopus Google Scholar). that both ceramide and AA in and their in the of or translocation of we showed that ceramide εPKC from the cytoplasm to the region and region the Golgi complex by staining that AA the translocation of εPKC to the Golgi complex Y. K. K. J. Biol. PubMed Scopus Google Scholar). 10 ceramide and AA were used to translocation of PKCs and the translocation to the Golgi complex at AA and The of that of that εPKC translocated to the Golgi complex ceramide generated by with in the of ceramide and that εPKC in the Golgi complex in the that translocation of εPKC to the Golgi complex both ceramide and AA translocated εPKC from the cytoplasm to the Golgi complex, the of was but in εPKC was in the Golgi complex with in the upon AA treatment εPKC in a the with fluorescence in the after ceramide or AA differences in the of εPKC from the Golgi The ceramide-stimulated of εPKC with the Golgi complex was by the from the Golgi complex to the the of εPKC was to translocated by distinct of εPKC with the Golgi The that, in fluorescence recovery in the Golgi complex was with fluorescence the εPKC with the staining in the of the Golgi complex that of εPKC in the Golgi complex in response to ceramide. the after AA the recovery from of the Golgi complex the that AA a of εPKC with the Golgi complex in with εPKC εPKC was to by ceramide but AA the that εPKC to but the εPKC translocated by AA. differences that distinct mechanisms are in the translocation of εPKC by ceramide and AA. different of ceramide and AA on PKC translocation for T. J. Biol. PubMed Scopus Google Scholar). with AA the translocation of to the but with ceramide had no effect on the the of εPKC for and εPKC we a of mutants and their translocation of the translocation in response to ceramide or AA with of the C1B domain the mutants to both ceramide and AA. that the C1B domain for the translocation by both ceramide and AA. the mechanisms the distinct translocation are differences in specific that the but the domain, in the and translocation that and C1B different in translocation. in the of and C1B was ceramide and AA, translocation of both and but the mutants both and C1B and were to that or C1B translocation. the translocation of was that of with the that the C1B domain of εPKC for phorbol the domain K. K. Y. Kikkawa U. J. Scopus Google Scholar). distinct of and C1B domain in the of J. Mol. 1995; 12: PubMed Scopus Google Scholar, J. Google used mutants of or C1B and showed no differences in that or the K. PubMed Scopus Google that translocation of δPKC by the C1B domain but and C1B of for the translocation. the was translocated to the with the or upon treatment of with or of T. J. Biol. PubMed Scopus Google Scholar). Thus, a of with that and C1B PKC used point in the and and C1B and to that the C1B domain responsible for the or and that the or the C1B domain for the translocation. The mutants a for and the to S. J. Biol. 1995; PubMed Scopus Google Scholar). Ceramide and AA the but the C1B that the εC1B domain for ceramide- and a only one of domain, in or for εPKC, ceramide the translocation of δPKC from the to the Golgi complex T. S. Y. Mol. Cell. Biol. PubMed Scopus Google Scholar). εPKC, δPKC to AA Y. K. J. Google Scholar). are the differences εPKC and that the εC1B for AA-induced translocation of the differences the C1B of εPKC and δPKC for their sensitivity to the of δPKC the C1B domain of εPKC, in response to AA, but the of εPKC the C1B domain of (ε(δC1B)), but translocated in response to AA. to a of the ε(δC1B) in response to ceramide the differences the and that εC1B domain to both ceramide and AA, but only to ceramide. differences in the C1B domain the of and δPKC to lipid second the differences εPKC and δPKC to the lipid second their AA to one of J. Nature. 1992; PubMed Scopus Google and AA by the of in the Nature. PubMed Scopus Google Scholar, PubMed Scopus Google Scholar, T. T. T. PubMed Scopus Google Scholar). εPKC activated with AA in the of δPKC activated with AA at K. Kikkawa U. J. 1995; PubMed Scopus Google Scholar). εPKC in and and in the Y. Ogita K. Res. PubMed Scopus Google Scholar). AA-induced of εPKC, but δPKC in in the of one with and in the of to T. Y. S. J. Biol. PubMed Scopus Google Scholar, Science. PubMed Scopus Google Scholar, Science. PubMed Scopus Google or T. J. Biol. PubMed Google Scholar). that ceramide are by or T. Y. S. J. Biol. PubMed Scopus Google that ceramide translocated and εPKC, α, to the in in T. Y. S. J. Biol. PubMed Scopus Google Scholar). that only but ceramide and AA the and of PKC to the in or PKC are in the subtype of PKC by specific activators and a in ceramide and AA εPKC to the Golgi complex by distinct mechanisms the C1B only or C1B domain for translocation. The differences in the C1B of εPKC and δPKC for their sensitivity to AA. that different of PKC translocation in response to different second and the distinct of the domain the to the The PKC 1The abbreviations used are: PKCprotein kinase CAAarachidonic acidFRAPfluorescent recovery after photobleachingGFPgreen fluorescent proteinεPKCε subtype of protein kinase CδPKCδ subtype of protein kinase CTPA12-O-tetradecanoylphorbol-13-acetateWGAwheat germ agglutininDGdiacylglycerolPSpseudosubstrateC1conserved region 1GFPgreen fluorescent proteinCHOChinese hamster ovaryPBSphosphate-buffered salinePDBuphorbol 12,13-dibutyrate 1The abbreviations used are: PKCprotein kinase CAAarachidonic acidFRAPfluorescent recovery after photobleachingGFPgreen fluorescent proteinεPKCε subtype of protein kinase CδPKCδ subtype of protein kinase CTPA12-O-tetradecanoylphorbol-13-acetateWGAwheat germ agglutininDGdiacylglycerolPSpseudosubstrateC1conserved region 1GFPgreen fluorescent proteinCHOChinese hamster ovaryPBSphosphate-buffered salinePDBuphorbol 12,13-dibutyrate family of serine/threonine protein kinase contains at least 10 subtypes. They are divided into three subgroups based on structural differences and requirement for activators (1Nishizuka Y. Nature. 1984; 308: 693-698Crossref PubMed Scopus (5740) Google Scholar, 2Nishizuka Y. Nature. 1988; 334: 661-665Crossref PubMed Scopus (3523) Google Scholar, 3Nishizuka Y. Science. 1992; 258: 607-614Crossref PubMed Scopus (4214) Google Scholar, 4Nishizuka Y. FASEB J. 1995; 9: 484-496Crossref PubMed Scopus (2352) Google Scholar). The conventional PKCs (cPKC; α, βI, βII, and γ) are Ca2+-dependent and activated by diacylglycerol or phorbol esters. The novel PKCs (nPKC; δ, ε, η, and θ) are activated by diacylglycerol (DG) or phorbol esters, but are Ca2+-independent (5Ono Y. Fujii T. Igarashi K. Kikkawa U. Ogita K. Nishizuka Y. Nucleic Acids Res. 1988; 16: 5199-5200Crossref PubMed Scopus (58) Google Scholar, 6Osada S. Mizuno K. Saido T.C. Suzuki K. Kuroki T. Ohno S. Mol. Cell. Biol. 1992; 12: 3930-3938Crossref PubMed Google Scholar, 7Hug J. PubMed Scopus Google Scholar). The PKCs and are to and are Ca2+-independent Y. Fujii T. Igarashi K. T. Kikkawa U. Nishizuka Y. U. S. PubMed Scopus Google Scholar, K. Mizuno K. S. S. S. Suzuki K. Ohno S. J. Biol. PubMed Google Scholar, Y. J. Biol. PubMed Google Scholar). protein kinase C arachidonic acid fluorescent recovery after fluorescent protein subtype of protein kinase C subtype of protein kinase C germ diacylglycerol region fluorescent protein hamster phorbol 12,13-dibutyrate protein kinase C arachidonic acid fluorescent recovery after fluorescent protein subtype of protein kinase C subtype of protein kinase C germ diacylglycerol region fluorescent protein hamster phorbol 12,13-dibutyrate PKCs an domain and a domain in the The domain of the PKCs contains a region a and a region The of εPKC to a of εPKC translocation J. Biol. PubMed Scopus Google Scholar, T. J. J. Biol. PubMed Scopus Google Scholar). the the in the of the domain, the by the The of activators to the domain a which the from the and the Biol. 9: PubMed Scopus Google Scholar). and phorbol to the domain Y. Nature. 1988; 334: 661-665Crossref PubMed Scopus (3523) Google Scholar, Y. Fujii T. Igarashi K. T. Kikkawa U. Nishizuka Y. U. S. PubMed Scopus Google Scholar, Biol. 1995; PubMed Scopus Google Scholar, J. Biol. PubMed Google Scholar). the domain that of εPKC J. Biol. PubMed Scopus Google Scholar, S. Mol. Biol. Cell. PubMed Scopus Google Scholar, Suzuki K. J. Y. T. J. Biol. PubMed Scopus Google Scholar). The domain of and and of and in a The C1B of and showed phorbol but for showed for phorbol K. K. Y. Kikkawa U. J. Scopus Google Scholar). or translocated to the in response to or phorbol translocation of C1B was only in phorbol T. J. Biol. PubMed Scopus Google Scholar). distinct for the and C1B in the of the J. Biol. PubMed Scopus Google Scholar). that the and C1B of PKCs are The of PKC only by and but by arachidonic acid (AA) K. Kikkawa U. J. 1995; PubMed Scopus Google Scholar, T. Y. K. Nishizuka Y. U. S. PubMed Scopus Google and ceramide J. PubMed Scopus Google Scholar, J. Res. PubMed Scopus Google Scholar). esters, lipid second induce translocation of and in that AA translocation of εPKC Y. J. Google and ceramide εPKC and δPKC from the to the cytoplasm T. Y. S. J. Biol. PubMed Scopus Google Scholar). Using fluorescent protein PKCs and we that AA εPKC, but from the cytoplasm to the Golgi complex Y. K. K. J. Biol. PubMed Scopus Google Scholar, Y. K. J. Google and that ceramide δPKC from the cytoplasm to the Golgi complex T. S. Y. Mol. Cell. Biol. PubMed Scopus Google Scholar). the mechanism we the of and δPKC that to ceramide and AA to the molecular mechanisms responsible for the translocation of of ceramide and AA lipid only to Ceramide in T. J. Biol. PubMed Google of Y. S. J. PubMed Google T. Y. S. J. Biol. PubMed Scopus Google Scholar, Science. PubMed Scopus Google Scholar, Science. PubMed Scopus Google and of PubMed Scopus Google Scholar). AA to a J. Nature. 1992; PubMed Scopus Google in the of a of T. Res. PubMed Scopus Google a of J. Res. PubMed Scopus Google Scholar, J. Biol. PubMed Scopus Google and to for J. Biol. PubMed Scopus Google Scholar). ceramide and AA the translocation of AA εPKC but δPKC in K. Kikkawa U. J. 1995; PubMed Scopus Google and the of J. Res. PubMed Scopus Google Scholar). S. J. Biol. PubMed Scopus Google that and δPKC translocated by of AA. T. J. Biol. PubMed Scopus Google that AA translocation of the domain of Ceramide δPKC and εPKC from the to the cytoplasm T. Y. S. J. Biol. PubMed Scopus Google and from the cytoplasm to the J. PubMed Scopus Google Scholar). that both ceramide and AA in and their in the of or translocation of we showed that ceramide εPKC from the cytoplasm to the region and region the Golgi complex by staining that AA the translocation of εPKC to the Golgi complex Y. K. K. J. Biol. PubMed Scopus Google Scholar). 10 ceramide and AA were used to translocation of PKCs and the translocation to the Golgi complex at AA and The of that of that εPKC translocated to the Golgi complex ceramide generated by with in the of ceramide and that εPKC in the Golgi complex in the that translocation of εPKC to the Golgi complex both ceramide and AA translocated εPKC from the cytoplasm to the Golgi complex, the of was but in εPKC was in the Golgi complex with in the upon AA treatment εPKC in a the with fluorescence in the after ceramide or AA differences in the of εPKC from the Golgi The ceramide-stimulated of εPKC with the Golgi complex was by the from the Golgi complex to the the of εPKC was to translocated by distinct of εPKC with the Golgi The that, in fluorescence recovery in the Golgi complex was with fluorescence the εPKC with the staining in the of the Golgi complex that of εPKC in the Golgi complex in response to ceramide. the after AA the recovery from of the Golgi complex the that AA a of εPKC with the Golgi complex in with εPKC εPKC was to by ceramide but AA the that εPKC to but the εPKC translocated by AA. differences that distinct mechanisms are in the translocation of εPKC by ceramide and AA. different of ceramide and AA on PKC translocation for T. J. Biol. PubMed Scopus Google Scholar). with AA the translocation of to the but with ceramide had no effect on the the of εPKC for and εPKC we a of mutants and their translocation of the translocation in response to ceramide or AA with of the C1B domain the mutants to both ceramide and AA. that the C1B domain for the translocation by both ceramide and AA. the mechanisms the distinct translocation are differences in specific that the but the domain, in the and translocation that and C1B different in translocation. in the of and C1B was ceramide and AA, translocation of both and but the mutants both and C1B and were to that or C1B translocation. the translocation of was that of with the that the C1B domain of εPKC for phorbol the domain K. K. Y. Kikkawa U. J. Scopus Google Scholar). distinct of and C1B domain in the of J. Mol. 1995; 12: PubMed Scopus Google Scholar, J. Google used mutants of or C1B and showed no differences in that or the K. PubMed Scopus Google that translocation of δPKC by the C1B domain but and C1B of for the translocation. the was translocated to the with the or upon treatment of with or of T. J. Biol. PubMed Scopus Google Scholar). Thus, a of with that and C1B PKC used point in the and and C1B and to that the C1B domain responsible for the or and that the or the C1B domain for the translocation. The mutants a for and the to S. J. Biol. 1995; PubMed Scopus Google Scholar). Ceramide and AA the but the C1B that the εC1B domain for ceramide- and a only one of domain, in or for εPKC, ceramide the translocation of δPKC from the to the Golgi complex T. S. Y. Mol. Cell. Biol. PubMed Scopus Google Scholar). εPKC, δPKC to AA Y. K. J. Google Scholar). are the differences εPKC and that the εC1B for AA-induced translocation of the differences the C1B of εPKC and δPKC for their sensitivity to the of δPKC the C1B domain of εPKC, in response to AA, but the of εPKC the C1B domain of (ε(δC1B)), but translocated in response to AA. to a of the ε(δC1B) in response to ceramide the differences the and that εC1B domain to both ceramide and AA, but only to ceramide. differences in the C1B domain the of and δPKC to lipid second the differences εPKC and δPKC to the lipid second their AA to one of J. Nature. 1992; PubMed Scopus Google and AA by the of in the Nature. PubMed Scopus Google Scholar, PubMed Scopus Google Scholar, T. T. T. PubMed Scopus Google Scholar). εPKC activated with AA in the of δPKC activated with AA at K. Kikkawa U. J. 1995; PubMed Scopus Google Scholar). εPKC in and and in the Y. Ogita K. Res. PubMed Scopus Google Scholar). AA-induced of εPKC, but δPKC in in the of one with and in the of to T. Y. S. J. Biol. PubMed Scopus Google Scholar, Science. PubMed Scopus Google Scholar, Science. PubMed Scopus Google or T. J. Biol. PubMed Google Scholar). that ceramide are by or T. Y. S. J. Biol. PubMed Scopus Google that ceramide translocated and εPKC, α, to the in in T. Y. S. J. Biol. PubMed Scopus Google Scholar). that only but ceramide and AA the and of PKC to the in or PKC are in the subtype of PKC by specific activators and a in ceramide and AA εPKC to the Golgi complex by distinct mechanisms the C1B only or C1B domain for translocation. The differences in the C1B of εPKC and δPKC for their sensitivity to AA. that different of PKC translocation in response to different second and the distinct of the domain the to the The of ceramide and AA lipid only to Ceramide in T. J. Biol. PubMed Google of Y. S. J. PubMed Google T. Y. S. J. Biol. PubMed Scopus Google Scholar, Science. PubMed Scopus Google Scholar, Science. PubMed Scopus Google and of PubMed Scopus Google Scholar). AA to a J. Nature. 1992; PubMed Scopus Google in the of a of T. Res. PubMed Scopus Google a of J. Res. PubMed Scopus Google Scholar, J. Biol. PubMed Scopus Google and to for J. Biol. PubMed Scopus Google Scholar). ceramide and AA the translocation of AA εPKC but δPKC in K. Kikkawa U. J. 1995; PubMed Scopus Google and the of J. Res. PubMed Scopus Google Scholar). S. J. Biol. PubMed Scopus Google that and δPKC translocated by of AA. T. J. Biol. PubMed Scopus Google that AA translocation of the domain of Ceramide δPKC and εPKC from the to the cytoplasm T. Y. S. J. Biol. PubMed Scopus Google and from the cytoplasm to the J. PubMed Scopus Google Scholar). that both ceramide and AA in and their in the of or translocation of we showed that ceramide εPKC from the cytoplasm to the region and region the Golgi complex by staining that AA the translocation of εPKC to the Golgi complex Y. K. K. J. Biol. PubMed Scopus Google Scholar). 10 ceramide and AA were used to translocation of PKCs and the translocation to the Golgi complex at AA and The of that of that εPKC translocated to the Golgi complex ceramide generated by with in the of ceramide and that εPKC in the Golgi complex in the that translocation of εPKC to the Golgi complex both ceramide and AA translocated εPKC from the cytoplasm to the Golgi complex, the of was but in εPKC was in the Golgi complex with in the upon AA treatment εPKC in a the with fluorescence in the after ceramide or AA differences in the of εPKC from the Golgi The ceramide-stimulated of εPKC with the Golgi complex was by the from the Golgi complex to the the of εPKC was to translocated by distinct of εPKC with the Golgi The that, in fluorescence recovery in the Golgi complex was with fluorescence the εPKC with the staining in the of the Golgi complex that of εPKC in the Golgi complex in response to ceramide. the after AA the recovery from of the Golgi complex the that AA a of εPKC with the Golgi complex in with εPKC εPKC was to by ceramide but AA the that εPKC to but the εPKC translocated by AA. differences that distinct mechanisms are in the translocation of εPKC by ceramide and AA. different of ceramide and AA on PKC translocation for T. J. Biol. PubMed Scopus Google Scholar). with AA the translocation of to the but with ceramide had no effect on the translocation. the of εPKC for and εPKC we a of mutants and their translocation of the translocation in response to ceramide or AA with of the C1B domain the mutants to both ceramide and AA. that the C1B domain for the translocation by both ceramide and AA. the mechanisms the distinct translocation are differences in specific The that the but the domain, in the and translocation that and C1B different in translocation. in the of and C1B was ceramide and AA, translocation of both and but the mutants both and C1B and were to that or C1B translocation. the translocation of was that of with the that the C1B domain of εPKC for phorbol the domain K. K. Y. Kikkawa U. J. Scopus Google Scholar). distinct of and C1B domain in the of J. Mol. 1995; 12: PubMed Scopus Google Scholar, J. Google used mutants of or C1B and showed no differences in that or the K. PubMed Scopus Google that translocation of δPKC by the C1B domain but and C1B of for the translocation. the was translocated to the with the or upon treatment of with or of T. J. Biol. PubMed Scopus Google Scholar). Thus, a of with that and C1B PKC translocation. used point in the and and C1B and to that the C1B domain responsible for the or and that the or the C1B domain for the translocation. The mutants a for and the to S. J. Biol. 1995; PubMed Scopus Google Scholar). Ceramide and AA the but the C1B that the εC1B domain for ceramide- and a only one of domain, in or for localization. εPKC, ceramide the translocation of δPKC from the to the Golgi complex T. S. Y. Mol. Cell. Biol. PubMed Scopus Google Scholar). εPKC, δPKC to AA Y. K. J. Google Scholar). are the differences εPKC and that the εC1B for AA-induced translocation of the differences the C1B of εPKC and δPKC for their sensitivity to the of δPKC the C1B domain of εPKC, in response to AA, but the of εPKC the C1B domain of (ε(δC1B)), but translocated in response to AA. to a of the ε(δC1B) in response to ceramide the differences the and that εC1B domain to both ceramide and AA, but only to ceramide. differences in the C1B domain the of and δPKC to lipid second messengers. the differences εPKC and δPKC to the lipid second their AA to one of J. Nature. 1992; PubMed Scopus Google and AA by the of in the Nature. PubMed Scopus Google Scholar, PubMed Scopus Google Scholar, T. T. T. PubMed Scopus Google Scholar). εPKC activated with AA in the of δPKC activated with AA at K. Kikkawa U. J. 1995; PubMed Scopus Google Scholar). εPKC in and and in the Y. Ogita K. Res. PubMed Scopus Google Scholar). AA-induced of εPKC, but δPKC in in the of one with and in the of to T. Y. S. J. Biol. PubMed Scopus Google Scholar, Science. PubMed Scopus Google Scholar, Science. PubMed Scopus Google or T. J. Biol. PubMed Google Scholar). that ceramide are by or T. Y. S. J. Biol. PubMed Scopus Google that ceramide translocated and εPKC, α, to the in in T. Y. S. J. Biol. PubMed Scopus Google Scholar). that only but ceramide and AA the and of PKC to the in or PKC are in the subtype of PKC by specific activators and a in ceramide and AA εPKC to the Golgi complex by distinct mechanisms the C1B only or C1B domain for translocation. The differences in the C1B of εPKC and δPKC for their sensitivity to AA. that different of PKC translocation in response to different second and the distinct of the domain the to the of The for of