Abstract
We investigated the effect of insulin on the expression of the enhancer of split- and hairy-related protein-2 gene in 3T3-L1 adipocytes and L6 myotubes. The level of enhancer of split- and hairy-related protein-2 mRNA was increased by insulin in both cells. While both wortmannin and LY294002 blocked the increase in 3T3-L1 adipocytes, and only PD98059 was effective in L6 myotubes. Although the increase by insulin in these cells was inhibited by treatment with actinomycin D, this was enhanced by treatment with cycloheximide. Furthermore, cyclic AMP increased the level of enhancer of split- and hairy-related protein-2 mRNA in both cells in an additive manner. Thus, we conclude that insulin and cyclic AMP induce the expression of the enhancer of split- and hairy-related protein-2 gene in both 3T3-L1 adipocytes and L6 myotubes, and that the gene expression enhanced by insulin is regulated by the cell type-specific pathway. The former requires a phosphoinositide 3-kinase pathway and the latter a mitogen-activated protein kinase pathway.
Key words
Insulin · Signaling pathway · Gene expression · Phosphoinositide 3-kinase · Mitogen-activated protein kinase
References
1
Massari M E, Murre C.
Helix-loop-helix proteins: regulators of transcription in eucaryotic organisms.
Mol Cell Biol.
2000;
20
429-440
2
Rossner M J, Dorr J, Gass P, Schwab M H, Nave K A.
SHARPs: mammalian enhancer-of-split- and hairy-related proteins coupled to neuronal stimulation.
Mol Cell Neurosci.
1997;
9
460-475
3
Shen M, Kawamoto T, Yan W, Nakamasu K, Tamagami M, Koyano Y, Noshiro M, Kato Y.
Molecular characterization of the novel basic helix-loop-helix protein DEC1 expressed in differentiated human embryo chondrocytes.
Biochem Biophys Res Comm.
1997;
236
294-298
4
Boudjelal M, Taneja R, Matsubara S, Bouillet P, Dolle P, Chambon P.
Overexpression of Stra13, a novel retinoic acid-inducible gene of the basic helix-loop-helix family, inhibits mesodermal and promotes neuronal differentiation of P19 cells.
Genes Dev.
1997;
11
2052-2065
5
Sun H, Teneja R.
Stra13 expression is associated with growth arrest and represses transcription through histone deacetylase (HDAC)-dependent and HDAC-independent mechanisms.
Proc Natl Acad Sci USA.
2000;
97
4058-4063
6
Zawel L, Yu J, Torrance C, Markowitz S, Kinzler K W, Vogelstein B, Zhou S.
DEC1 is a downstream target of TGF-beta with sequence-specific transcriptional repressor activities.
Proc Natl Acad Sci U S A.
2002;
99
2848-2853
7
Yamada K, Kawata H, Mizutani T, Arima T, Yazawa T, Matsuura K, Shou Z, Sekiguchi T, Yoshino M, Kajitani T, Miyamoto K.
Gene expression of basic helix-loop-helix transcription factor, SHARP-2, is regulated by gonadotropins in the rat ovary and MA-10 cells.
Biol Reprod.
2004;
70
76-82
8
Inova A V, Inova S V, Danilkovitch-Miagkova A, Lerman M I.
Regulation of STRA13 by the von Hippel-Lindau tumor supressor protein, hypoxia, and the UBC9/ubiquitin proteasome degradation pathway.
J Biol Chem.
2001;
276
15 306-15 315
9
Yun Z, Maecker H L, Johnson R S, Giaccia A J.
Inhibition of PPAR gamma 2 gene expression by the HIF-1-regulated gene DEC1/Stra13: a mechanism for regulation of adipogenesis by hypoxia.
Dev Cell.
2002;
2
331-341
10
Honma S, Kawamoto T, Takagi Y, Fujimoto K, Sato F, Noshiro M, Kato Y, Honma K.
Dec1 and Dec2 are regulators of the mammalian molecular clock.
Nature.
2002;
419
841-844
11
Sun H, Lu B, Li R-Q, Flavell R A, Taneja R.
Defective T cell activation and autoimmune disorder in Stra13-deficient mice.
Nat Immunol.
2001;
2
1040-1047
12
Yamada K, Kawata H, Shou Z, Mizutani T, Noguchi T, Miyamoto K.
Insulin induces the expression of the SHARP-2/Stra13LDEC1 gene via a phosphoinositide 3-kinase pathway.
J Biol Chem.
2003;
278
30719-30724
13
Asai Y, Yamada K, Watanabe T, Keng V W, Noguchi T.
Insulin stimulates expression of pyruvate kinase M gene in 3T3-L1 adipocytes.
Biosci Biotech Bioch.
2003;
67
1272-1277
14
Printz R L, Koch S, Potter L R, O’Doherty R M, Tiesinga J J, Moritz S, Granner D K.
Hexokinase II mRNA and gene structure, regulation by insulin, and evolution.
J Biol Chem.
1993;
268
5209-5219
15
Bates S H, Gardiner J V, Jones R B, Bloom S R, Bailey C J.
Acute stimulation of glucose uptake by leptin in L6 muscle cells.
Horm Metab Res.
2002;
34
111-115
16
Shou Z, Yamada K, Inazu T, Kawata H, Hirano S, Mizutani T, Yazawa T, Sekiguchi T, Yoshino M, Kajitani T, Miyamoto K.
Genomic structure and promoter analysis of the mouse zinc-fingers and homeoboxes 1 (ZHX1) gene.
Gene.
2003;
302
83-94
17
Agati J M, Yeagley D, Quinn P G.
Assessment of the roles of mitogen-activated protein kinase, phosphatidylinositol 3-kinase, protein kinase B, and protein kinase C in insulin inhibition of cAMP-induced phosphoenolpyruvate carboxykinase gene transcription.
J Biol Chem.
1998;
273
18751-18759
18
Tortorella L L, Milasincic D J, Pilch P F.
Critical proliferation-independent window for basic fibroblast growth factor repression of myogenesis via the p42/p44 MAPK signaling pathway.
J Biol Chem.
2001;
276
13709-13717
19
Haruta T, Urio T, Kawahara J, Takano A, Egawa K, Sharma P M, Olefsky J M, Kobayashi M.
A rapamycin-sensitive pathway down-regulates insulin signaling via phosphorylation and proteasomal degradation of insulin receptor substrate-1.
Mol Endocrinol.
2000;
14
783-794
20
Pirola L, Bonnafous S, Johnston A M, Chaussade C, Portis F, van Obberghen E.
Phosphoinositide 3-kinase-mediated reduction of insulin receptor substrate-1/2 protein expression via different mechanisms contributes to the insulin-induced desensitization of its signaling pathways in L6 muscle cells.
J Biol Chem.
2003;
278
15641-15651
21
Fasshauer M, Klein J, Lossner U, Paschke R.
Interleukin (IL)-6 mRNA expression is stimulated by insulin, isoproterenol, tumour necrosis factor alpha, growth hormone, and IL-6 in 3T3-L1 adipocytes.
Horm Metab Res.
2003;
35
147-152
22
Hirano S, Yamada K, Kawata H, Shou Z, Mizutani T, Shigematsu Y, Mayumi M, Miyamoto K.
The rat enhancer of split- and hairy-relatedprotein-2 gene: hepatic expression, genomic structure, and promoter analysis.
Arch Biochem Biophys.
2004;
422
81-90
23
Teramoto M, Nakamasu K, Noshiro M, Matsuda Y, Gotoh O, Shen M, Tsutsumi S, Kawamoto T, Iwamoto Y, Kato Y.
Gene structure and chromosomal location of a human bHLH transcriptional factor DEC1 × Stra13 × SHARP-2/BHLHB2.
J Biochem.
2001;
129
391-396
24
Osawa H, Whitesell R L, Printz R L, Granner D K.
Regulation of hexokinase II gene transcription and glucose phosphorylation by catecholamines, cyclic AMP, and insulin.
Diabetes.
1995;
44
1426-1432
Dr. K. Yamada
Department of Biochemistry, Faculty of Medical Sciences
University of Fukui and CREST, JST · Fukui 910-1193 · Japan
Phone: +81(776)61-8316
Fax: +81(776)61-8102 ·
Email: kazuya@fmsrsa.fukui-med.ac.jp.