title: Developmentally spliced PKCbetaII provides a possible link between mTORC2 and Akt kinase to regulate 3T3-L1 adipocyte insulin-stimulated glucose transport.
abstract: Functional adipocyte glucose disposal is a key component of global glucose homeostasis. PKCbetaII is involved in rat skeletal muscle cell ISGT. Western blot analysis and real-time PCR revealed 3T3-L1 cells developmentally regulated PKCbeta splicing such that PKCbetaI was downregulated and PKCbetaII was upregulated during the course of differentiation. An initial glucose uptake screen using PKC inhibitor LY379196 pointed to a PKC isozyme other than PKCzeta mediating 3T3-L1 adipocyte ISGT. Subsequent use of PKCbetaII inhibitor CGP53353 pointed to a role for PKCbetaII in ISGT. Western blot analysis showed that CGP53353 specifically inhibited phosphorylation of PKCbetaII Serine 660. Subcellular fractionation and immunofluorescence demonstrated that PKCbetaII regulates GLUT4 translocation. Further Western blot, immunofluorescence and co-immunoprecipitation analysis reveal that PKCbetaII inhibition does not affect mTORC2 activity yet abrogates phosphorylation of Akt Serine 473. PKCbetaII regulates GLUT4 translocation by regulating Akt phosphorylation and thus activity.
authors.affiliation: Department of Molecular Medicine, College of Medicine, University of South Florida, The Research Service, James A. Haley Veterans Hospital, Tampa, FL 33612, USA.
authors.forename: E
authors.initials: E
authors.lastname: Kleiman
authors.forename: G
authors.initials: G
authors.lastname: Carter
authors.forename: T
authors.initials: T
authors.lastname: Ghansah
authors.forename: N A
authors.initials: NA
authors.lastname: Patel
authors.forename: D R
authors.initials: DR
authors.lastname: Cooper
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000378:metabolism*; D017398:Alternative Splicing; D000818:Animals; D003847:Deoxyglucose / Q000378:metabolism; D005947:Glucose / Q000378:metabolism*; D051275:Glucose Transporter Type 4 / Q000378:metabolism*; D007328:Insulin / Q000378:metabolism* / Q000494:pharmacology; D008698:Mesylates / Q000494:pharmacology; D051379:Mice; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D064546:Protein Kinase C beta; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D011758:Pyrroles / Q000494:pharmacology; D051381:Rats; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors
chemical_list: C434605:5,21 - 12,17-dimetheneo-18H-dibenzo(i,o)pyrrolo(3,4-1)(1,8)diazacyclohexandecine-18,10(19H)dione,8((dimethylamino)methyl)-6,7,8,9,10,11-hexahydro,monomethanesulfonate; C504923:Crtc2 protein, mouse; D051275:Glucose Transporter Type 4; D007328:Insulin; D008698:Mesylates; D047428:Protein Kinase Inhibitors; D011758:Pyrroles; C495693:Slc2a4 protein, mouse; D015534:Trans-Activators; D014157:Transcription Factors; D003847:Deoxyglucose; D051057:Proto-Oncogene Proteins c-akt; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D005947:Glucose
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