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Dominant negative α-subunit of farnesyl- and geranylgeranyl-transferase I inhibits insulin-induced differentiation of 3T3-L1 pre-adipocytes

International Journal of Obesity volume 27, pages 40–47 (2003)Cite this article

Abstract

OBJECTIVE: To investigate whether the expression of a dominant negative (DN) farnesyl- and geranygeranyl-transferase I (FTase/GGTase I) α-subunit in 3T3-L1 pre-adipocytes can inhibit insulin's ability to induce differentiation.

DESIGN: 3T3-L1 pre-adipocytes were stably transfected with vector alone or vector expressing a mutated DN FTase/GGTase I α-subunit (S60A)(S62A) and incubated in serum-free medium in the absence and presence of insulin.

MEASUREMENTS: Various assays were performed to determine the effect of DN FTase/GGTase I α-subunit expression in 3T3-L1 pre-adipocyte on insulin-induced DNA synthesis, cell count, phosphorylation of the FTase/GGTase I α-subunit, FTase and GGTase I activity, amounts of prenylated p21Ras and RhoA, phosphorylation of MAP kinase and Akt, and differentiation to mature fat cells.

RESULTS: Expression of DN FTase/GGTase I α-subunit inhibited insulin's ability to increase DNA synthesis, cell count, FTase and GGTase I activity, amounts of prenylated p21Ras and RhoA, and magnitude of phosphorylation of MAP kinase. Expression of DN FTase/GGTase I α-subunit in 3T3-L1 pre-adipocytes was without effect on insulin-induced Akt phosphorylation.

CONCLUSION: Expression of DN FTase/GGTase I α-subunit inhibits insulin-induced differentiation of 3T3-L1 pre-adipocytes to mature adipocytes, and thus could indicate potential therapeutic avenues to assuage the deleterious effects of obesity and type 2 diabetes.

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Acknowledgements

This work was supported by the Research Service of the Department of Veterans Affairs (to MLG). Dr Goalstone is a recipient of the VA Career Development Award.

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  1. Research Service, Denver VA Medical Center, Denver, Colorado, USA

    C S Solomon, J W Leitner & M L Goalstone

  2. Department of Medicine, University of Colorado Health Sciences Center, Denver, Colorado, USA

    M L Goalstone

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Correspondence to M L Goalstone.

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Solomon, C., Leitner, J. & Goalstone, M. Dominant negative α-subunit of farnesyl- and geranylgeranyl-transferase I inhibits insulin-induced differentiation of 3T3-L1 pre-adipocytes. Int J Obes 27, 40–47 (2003). https://doi.org/10.1038/sj.ijo.0802189

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