Developmentally spliced PKCbetaII provides a possible link between mTORC2 and Akt kinase to regulate 3T3-L1 adipocyte insulin-stimulated glucose transport.
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.
Kleiman
E
E
Department of Molecular Medicine, College of Medicine, University of South Florida, The Research Service, James A. Haley Veterans Hospital, Tampa, FL 33612, USA.
Carter
G
G
Ghansah
T
T
Patel
N A
NA
Cooper
D R
DR
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
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
J Cell Physiol. 1996 Apr;167(1):113-20
8698828
Mol Endocrinol. 2005 Apr;19(4):1067-77
15650020
Nature. 2001 Feb 8;409(6821):729-33
11217863
Gene. 2000 Jun 13;251(1):45-53
10863095
J Cell Biochem. 2003 Oct 15;90(3):561-72
14523990
Mol Genet Metab. 2008 Feb;93(2):210-5
17950644
J Biol Chem. 2005 Dec 9;280(49):40642-9
16216880
J Lipid Res. 2001 Jun;42(6):910-5
11369798
Endocr Rev. 2004 Apr;25(2):177-204
15082519
Nature. 2006 Dec 14;444(7121):847-53
17167472
Mol Endocrinol. 1996 Oct;10(10):1273-81
9121494
EMBO J. 2008 Jul 23;27(14):1919-31
18566587
Methods Mol Biol. 2008;456:307-15
18516571
Endocrinology. 2003 Sep;144(9):3765-73
12933646
J Mol Biol. 2003 May 9;328(4):805-19
12729756
Methods Mol Med. 2003;83:105-11
12619721
Nat Rev Mol Cell Biol. 2002 Apr;3(4):267-77
11994746
Arch Biochem Biophys. 1999 Dec 1;372(1):69-79
10562418
Gene. 2005 Mar 28;348:157-65
15777692
Clin Cancer Res. 1995 Aug;1(8):813-21
9816050
J Biol Chem. 2004 Nov 12;279(46):47720-5
15364915
Am J Physiol Endocrinol Metab. 2005 Oct;289(4):E551-61
15928024
Mol Cell Biol. 1997 May;17(5):2425-35
9111311
Mol Biol Cell. 2008 Oct;19(10):4032-41
18653467
Diabetes. 1999 Oct;48(10):1922-9
10512355
Mol Endocrinol. 2004 Apr;18(4):899-911
14752056
Diabetologia. 2009 May;52(5):901-11
19252893
Physiol Rev. 2008 Oct;88(4):1341-78
18923184
J Lipid Res. 2009 Apr;50 Suppl:S266-71
19033211
Nat Genet. 2008 Jan;40(1):51-60
17994017
Invest Ophthalmol Vis Sci. 2000 Jun;41(7):1876-84
10845612
J Biol Chem. 2006 Jul 14;281(28):19145-55
16698794
J Biol Chem. 2001 Jun 22;276(25):22648-54
11283022
Cancer Res. 2009 Mar 1;69(5):1821-7
19244117