Ectopic Expression of Protein Kinase CβII, -δ, and -ϵ, but Not -βI or -ζ, Provide for Insulin Stimulation of Glucose Uptake in NIH-3T3 Cells

1999 ◽  
Vol 372 (1) ◽  
pp. 69-79 ◽  
Author(s):  
Denise R. Cooper ◽  
James E. Watson ◽  
Niketa Patel ◽  
Philip Illingworth ◽  
Mildred Acevedo-Duncan ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Glucose Uptake ◽  
Ectopic Expression ◽  
3T3 Cells ◽  
Insulin Stimulation ◽  
Nih 3T3 ◽  
Stimulation Of ◽  
Nih 3T3 Cells

Ectopic Expression of Protein Kinase CβII, -δ, and -ϵ, but Not -βI or -ζ, Provide for Insulin Stimulation of Glucose Uptake in NIH-3T3 Cells

2001 ◽  
Vol 388 (1) ◽  
pp. 178
Author(s):  
Denise R. Cooper ◽  
James E. Watson ◽  
Niketa Patel ◽  
Philip Illingworth ◽  
Mildred Acevedo-Duncan ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Glucose Uptake ◽  
Ectopic Expression ◽  
3T3 Cells ◽  
Insulin Stimulation ◽  
Nih 3T3 ◽  
Stimulation Of ◽  
Nih 3T3 Cells

scholarly journals Glut4 Storage Vesicles without Glut4: Transcriptional Regulation of Insulin-Dependent Vesicular Traffic

2004 ◽  
Vol 24 (16) ◽  
pp. 7151-7162 ◽  
Author(s):  
Danielle N. Gross ◽  
Stephen R. Farmer ◽  
Paul F. Pilch
Keyword(s):  
Insulin Receptor ◽  
Glucose Uptake ◽  
Glucose Transporter ◽  
Ectopic Expression ◽  
3T3 Cells ◽  
Vesicular Traffic ◽  
Insulin Dependent ◽  
Nih 3T3 ◽  
Vesicular Compartment ◽  
Nih 3T3 Cells

ABSTRACT Two families of transcription factors that play a major role in the development of adipocytes are the CCAAT/enhancer-binding proteins (C/EBPs) and the peroxisome proliferator-activated receptors (PPARs), in particular PPARγ. Ectopic expression of either C/EBPα or PPARγ in NIH 3T3 fibroblasts results in the conversion of these cells to adipocyte-like cells replete with fat droplets. NIH 3T3 cells ectopically expressing C/EBPα (NIH-C/EBPα) differentiate into adipocytes and exhibit insulin-stimulated glucose uptake, whereas NIH 3T3 cells ectopically expressing PPARγ (NIH-PPARγ) differentiate but do not exhibit any insulin-stimulated glucose uptake, nor do they express any C/EBPα. The reason for the lack of insulin-responsive glucose uptake in the NIH-PPARγ cells is their virtual lack of the insulin-responsive glucose transporter, Glut4. The NIH-PPARγ cells express functionally active components of the insulin receptor-signaling pathway (the insulin receptor, IRS-1, phosphatidylinositol 3-kinase, and Akt2) at levels comparable to those in responsive cell lines. They also express components of the insulin-sensitive vesicular transport machinery, namely, VAMP2, syntaxin-4, and IRAP, the last of these being the other marker of insulin-regulated vesicular traffic along with Glut4. Interestingly, the NIH-PPARγ cells show normal insulin-dependent translocation of IRAP and form an insulin-responsive vesicular compartment as assessed by cell surface biotinylation and sucrose velocity gradient analysis, respectively. Moreover, expression of a Glut4-myc construct in the NIH-PPARγ cells results in its insulin-dependent translocation to the plasma membrane as assessed by immunofluorescence and Western blot analysis. Based on these data, we conclude that major role of C/EBPα in the context of the NIH-PPARγ cells is to regulate Glut4 expression. The differentiated cells possess a large insulin-sensitive vesicular compartment with negligible Glut4, and Glut4 translocation can be reconstituted on expression of this transporter.

scholarly journals Both the Catalytic and Regulatory Domains of Protein Kinase C Chimeras Modulate the Proliferative Properties of NIH 3T3 Cells

1997 ◽  
Vol 272 (45) ◽  
pp. 28793-28799 ◽  
Author(s):  
Péter Ács ◽  
Qiming J. Wang ◽  
Krisztina Bögi ◽  
Adriana M. Marquez ◽  
Patricia S. Lorenzo ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
3T3 Cells ◽  
Regulatory Domains ◽  
Kinase C ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

Mutation of amino acids in pp60c-src that are phosphorylated by protein kinases C and A

1989 ◽  
Vol 9 (6) ◽  
pp. 2453-2463
Author(s):  
P Yaciuk ◽  
J K Choi ◽  
D Shalloway
Keyword(s):  
Protein Kinase ◽  
3T3 Cells ◽  
Triple Mutant ◽  
Dependent Protein Kinase ◽  
Mutant Proteins ◽  
Tetradecanoyl Phorbol Acetate ◽  
Dependent Protein ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

The product of the c-src proto-oncogene, pp60c-src, is phosphorylated at Ser-17 by cyclic AMP-dependent protein kinase A and at Ser-12 by calcium-phospholipid-dependent protein kinase C (when stimulated by 12-O-tetradecanoyl phorbol acetate). We tested the effects of Ser----Ala and Ser----Glu mutations at these sites in pp60c-src and in pp60c-src(F527) (a mutant whose transforming activities are enhanced by Tyr-527----Phe mutation) by transfecting single-, double-, and triple-mutant src expression plasmids into NIH 3T3 cells. Tryptic phosphopeptide analyses of the mutant proteins confirmed prior biochemical identifications of the phosphorylation sites and showed that neither separate nor coordinate mutations at Ser-12 and Ser-17 affected Tyr-416, Tyr-527, or Ser-48 phosphorylation or prevented mitosis-specific phosphorylations of either pp60c-src or pp60c-src(F527). Ser-12 mutation did not affect phosphorylation of the Ser-17-containing peptide, but mutation of Ser-17 significantly increased phosphorylation at Ser-12. Specific kinase activities (both with and without in vivo 12-O-tetradecanoyl phorbol acetate treatment) and the abilities of pp60c-src and pp60c-src(F527) to induce foci, transformed morphologies, and anchorage-independent growth were unaffected by any of the serine mutations. Thus, pp60c-src transforming activity in NIH 3T3 cells is relatively insensitive to phosphorylation at these sites, but there is a suggestion that Ser-17 phosphorylation may have a subtle regulatory effect.

Stimulation of cell growth and proliferation in NIH-3T3 cells by onion and garlic oils

1986 ◽  
Vol 2 (3) ◽  
pp. 369-378 ◽  
Author(s):  
Judith T. Zelikoff ◽  
Norman M. Atkins ◽  
Sidney Belman
Keyword(s):  
Cell Growth ◽  
3T3 Cells ◽  
Cell Growth And Proliferation ◽  
Nih 3T3 ◽  
Stimulation Of ◽  
Nih 3T3 Cells

scholarly journals Cyclic AMP Blocks Cell Growth through Raf-1-Dependent and Raf-1-Independent Mechanisms

2002 ◽  
Vol 22 (11) ◽  
pp. 3717-3728 ◽  
Author(s):  
Nicolas Dumaz ◽  
Yvonne Light ◽  
Richard Marais
Keyword(s):  
Protein Kinase ◽  
Cell Growth ◽  
Cyclic Amp ◽  
3T3 Cells ◽  
Extracellular Signal ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

ABSTRACT It is widely accepted that cyclic AMP (cAMP) can block cell growth by phosphorylating Raf-1 on serine 43 and inhibiting signaling to extracellular signal-regulated protein kinase. We show that the suppression of Raf-1 by cAMP is considerably more complex than previously reported. When cellular cAMP is elevated, Raf-1 is phosphorylated on three residues (S43, S233, and S259), which work independently to block Raf-1. Both Ras-dependent and Ras-independent processes are disrupted. However, when cAMP-insensitive versions of Raf-1 are expressed in NIH 3T3 cells, their growth is still strongly suppressed when cAMP is elevated. Thus, although Raf-1 appears to be an important cAMP target, other pathways are also targeted by cAMP, providing alternative mechanisms that lead to suppression of cell growth.

scholarly journals Insulin directly stimulates mitochondrial glucose oxidation in the heart

2020 ◽  
Vol 19 (1) ◽  
Author(s):  
Qutuba G. Karwi ◽  
Cory S. Wagg ◽  
Tariq R. Altamimi ◽  
Golam M. Uddin ◽  
Kim L. Ho ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Glucose Uptake ◽  
Glucose Oxidation ◽  
Glycogen Synthase ◽  
Acid Oxidation ◽  
Mouse Heart ◽  
Insulin Stimulation ◽  
Direct Stimulation ◽  
Gsk 3Β ◽  
Stimulation Of

Abstract Background Glucose oxidation is a major contributor to myocardial energy production and its contribution is orchestrated by insulin. While insulin can increase glucose oxidation indirectly by enhancing glucose uptake and glycolysis, it also directly stimulates mitochondrial glucose oxidation, independent of increasing glucose uptake or glycolysis, through activating mitochondrial pyruvate dehydrogenase (PDH), the rate-limiting enzyme of glucose oxidation. However, how insulin directly stimulates PDH is not known. To determine this, we characterized the impacts of modifying mitochondrial insulin signaling kinases, namely protein kinase B (Akt), protein kinase C-delta (PKC-δ) and glycogen synthase kinase-3 beta (GSK-3β), on the direct insulin stimulation of glucose oxidation. Methods We employed an isolated working mouse heart model to measure the effect of insulin on cardiac glycolysis, glucose oxidation and fatty acid oxidation and how that could be affected when mitochondrial Akt, PKC-δ or GSK-3β is disturbed using pharmacological modulators. We also used differential centrifugation to isolate mitochondrial and cytosol fraction to examine the activity of Akt, PKC-δ and GSK-3β between these fractions. Data were analyzed using unpaired t-test and two-way ANOVA. Results Here we show that insulin-stimulated phosphorylation of mitochondrial Akt is a prerequisite for transducing insulin’s direct stimulation of glucose oxidation. Inhibition of mitochondrial Akt completely abolishes insulin-stimulated glucose oxidation, independent of glucose uptake or glycolysis. We also show a novel role of mitochondrial PKC-δ in modulating mitochondrial glucose oxidation. Inhibition of mitochondrial PKC-δ mimics insulin stimulation of glucose oxidation and mitochondrial Akt. We also demonstrate that inhibition of mitochondrial GSK3β phosphorylation does not influence insulin-stimulated glucose oxidation. Conclusion We identify, for the first time, insulin-stimulated mitochondrial Akt as a prerequisite transmitter of the insulin signal that directly stimulates cardiac glucose oxidation. These novel findings suggest that targeting mitochondrial Akt is a potential therapeutic approach to enhance cardiac insulin sensitivity in condition such as heart failure, diabetes and obesity.

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