Uncoupling protein 3 (UCP3) stimulates glucose uptake in muscle cells through a phosphoinositide 3-kinase-dependent mechanism

被引:75
作者
Huppertz, C
Fischer, BM
Kim, YB
Kotani, K
Vidal-Puig, A
Slieker, LJ
Sloop, KW
Lowell, BB
Kahn, BB
机构
[1] Beth Israel Deaconess Med Ctr, Diabet Unit, Dept Med, Div Endocrinol & Metab, Boston, MA 02215 USA
[2] Harvard Univ, Sch Med, Boston, MA 02215 USA
[3] Eli Lilly & Co, Lilly Res Labs, Endocrine Res Div, Indianapolis, IN 46285 USA
关键词
D O I
10.1074/jbc.M011708200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
UCP3 is a mitochondrial membrane protein expressed in humans selectively in skeletal muscle. To determine the mechanisms by which UCP3 plays a role in regulating glucose metabolism, we expressed human UCP3 in L6 myotubes by adenovirus-mediated gene transfer and in H9C2 cardiomyoblasts by stable transfection with a tetracycline-repressible UCP3 construct. Expression of UCP3 in L6 myotubes increased 2-deoxyglucose uptake S-fold and cell surface GLUT4 2.3-fold, thereby reaching maximally insulin-stimulated levels in control myotubes. Wortmannin, LY 294002, or the tyrosine kinase inhibitor genistein abolished the effect of UCP3 on glucose uptake, and wortmannin inhibited UCP3-induced GLUT4 cell surface recruitment. UCP3 overexpression increased phosphotyrosine-associated phosphoinositide 3-kinase (PI3K) activity 2.2-fold compared with control cells (p < 0.05). UCP3 overexpression increased lactate release 1.5- to 2-fold above control cells, indicating increased glucose metabolism. In H9C2 cardiomyoblasts stably transfected with UCP3 under control of a tetracycline-repressible promotor, removal of doxycycline resulted in detectable levels of UCP3 at 12 h and 2.2-fold induction at 7 days compared with 12 h, In parallel, glucose transport increased 1.3- and 8-fold at 12 h and 7 days, respectively, and the stimulation was inhibited by wortmannin or genistein, p85 association with membranes was increased 5.5-fold and phosphotyrosine-associated PI3K activity 3.8-fold. In contrast, overexpression of UCP3 in 3T3-L1 adipocytes did not alter glucose uptake, suggesting tissue-specific effects of human UCP3. Thus, UCP3 stimulates glucose transport and GLUT4 translocation to the cell surface in cardiac and skeletal muscle cells by activating a PI3K dependent pathway.
引用
收藏
页码:12520 / 12529
页数:10
相关论文
共 51 条
[31]  
MITSUMOTO Y, 1992, J BIOL CHEM, V267, P4957
[32]   Up-regulation of uncoupling protein 3 by thyroid hormone, peroxisome proliferator-activated receptor ligands and 9-cis retinoic acid in L6 myotubes [J].
Nagase, I ;
Yoshida, S ;
Canas, X ;
Irie, Y ;
Kimura, K ;
Yoshida, T ;
Saito, M .
FEBS LETTERS, 1999, 461 (03) :319-322
[33]   Role for uncoupling protein-2 as a regulator of mitochondrial hydrogen peroxide generation [J].
NegreSalvayre, A ;
Hirtz, C ;
Carrera, G ;
Cazenave, R ;
Troly, M ;
Salvayre, R ;
Penicaud, L ;
Casteilla, L .
FASEB JOURNAL, 1997, 11 (10) :809-815
[34]   Role for mitochondrial oxidants as regulators of cellular metabolism [J].
Nemoto, S ;
Takeda, K ;
Yu, ZX ;
Ferrans, VJ ;
Finkel, T .
MOLECULAR AND CELLULAR BIOLOGY, 2000, 20 (19) :7311-7318
[35]   Construction and characterization of a one-plasmid system for the controlled expression of genes in mammalian cells by tetracycline [J].
OBrien, K ;
Otto, K ;
Rao, RN .
GENE, 1997, 184 (01) :115-120
[36]  
OKADA T, 1994, J BIOL CHEM, V269, P3568
[37]   TYROSINE KINASE-DEFICIENT MUTANT HUMAN INSULIN-RECEPTORS (MET(1153)-]ILE) OVEREXPRESSED IN TRANSFECTED RAT ADIPOSE-CELLS FAIL TO MEDIATE TRANSLOCATION OF EPITOPE-TAGGED GLUT4 [J].
QUON, MJ ;
GUERREMILLO, M ;
ZARNOWSKI, MJ ;
BUTTE, AJ ;
EM, M ;
CUSHMAN, SW ;
TAYLOR, SI .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (12) :5587-5591
[38]   Human uncoupling proteins and obesity [J].
Schrauwen, P ;
Walder, K ;
Ravussin, E .
OBESITY RESEARCH, 1999, 7 (01) :97-105
[39]   Skeletal muscle uncoupling protein 3 expression is a determinant of energy expenditure in Pima Indians [J].
Schrauwen, P ;
Xia, J ;
Bogardus, C ;
Pratley, RE ;
Ravussin, E .
DIABETES, 1999, 48 (01) :146-149
[40]   The human uncoupling protein-3 gene - Genomic structure, chromosomal localization, and genetic basis for short and long form transcripts [J].
Solanes, G ;
VidalPuig, A ;
Grujic, D ;
Flier, JS ;
Lowell, BB .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (41) :25433-25436