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The inhibitory guanine nucleotide-binding protein Gi2α induces and potentiates adipocyte differentiation

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Lipids

Abstract

The present study further elucidates the involvement of the α-subunit of the GTP-binding protein Gi2 in the differentiation of murine 313-L1 cells. Control and vector-transfected cells attained a fully differentiated adipocyte phenotype showing ample lipid droplets. Cells expressing wild type (WT)-Gi2α or the constitutively active R179E-Gi2α, however, became enlarged, less confluent, and produced large amounts of lipids. Differentiation consistently increased the triglyceride (TAG) content in control cells. In both WT-Gi2α and R179E-Gi2α clones, a marked increase in TAG could be detected even prior to insulin/dexamethasone/isobutyl methylxanthine exposure. The activity of palmitoyl-CoA synthetase (PCS) and glycerophosphate acyltransferase (GPAT) also increased upon differentiation. WT-Gi2α and R179E-Gi2α overexpression also enhanced PCS and GPAT activities even before differentiation medium was added. The total amount of phospholipids (PL) generally increased upon differentiation; however, pre- and postdifferentiation values were insignificantly different in cells expressing WT-Gi2α and R179E-Gi2α. Differentiation altered the PL profile with a relative shift from phosphatidylcholine and phosphatidylethanolamine to phosphatidylinositol (PI) in differentiated cells. Finally, differentiation yielded a general increase in the activity of basal PI-phospholipase-C activity. Again, cells expressing WT-Gi2α and R179E-Gi2α demonstrated elevated enzyme activity and enhanced second messenger accumulation subsequent to differentiation. In summary, cells with the R179E-mutants of Gi2α exhibited stimulated lipid turnover and accumulation in both undifferentiated and differentiated cells.

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Abbreviations

DAG:

diacylglycerol

G204A-Gi2α:

constitutively inactivated (receptor dissociated) Gi2α mutant

GPAT:

glycerophosphate acyltransferase

GPDH:

glycerophosphate dehydrogenase

IGF:

insulin-like growth factor

IP:

1,4,5-inositol trisphosphate

MAPK:

mitogen-activating protein kinase

PBS:

phosphate-buffered saline

PCS:

palmitoyl-CoA synthetase

PI:

phosphatidylinositol

PIP2 :

phosphatidylinositol 4,5-bisphosphate

PI-PLC:

phosphatidylinositol phospholipase-C

PKC:

protein kinase C

PL:

phospholipid

pZipNeo:

pZipNeo(SX)V (retroviral vector)

R179E- and Q205L-Gi2α:

constitutively activated (GTPase deficient) Gi2α mutant

TAG:

triglyceride

WT:

wild type

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  1. Pengfei Wu (Visiting Scholar)

    Present address: Department of Biochemistry, Xinjiang Medical College, Xinjiang, China

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  1. Institute of Medical Biochemistry, University of Oslo, P.O. Box 1112, Blindern, 0317, Oslo, Norway

    Kjerstin E. Høvik, Pengfei Wu (Visiting Scholar) & Jan O. Gordeladze

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  1. Kjerstin E. Høvik
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Høvik, K.E., Wu, P. & Gordeladze, J.O. The inhibitory guanine nucleotide-binding protein Gi2α induces and potentiates adipocyte differentiation. Lipids 34, 355–362 (1999). https://doi.org/10.1007/s11745-999-0373-9

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