The insulin-sensitive glucose transporter, GLUT4, interacts physically with Daxx - Two proteins with capacity to bind Ubc9 and conjugated to SUMO1

被引:61
作者
Lalioti, VS [1 ]
Vergarajauregui, S [1 ]
Pulido, D [1 ]
Sandoval, IV [1 ]
机构
[1] Univ Autonoma Madrid, CSIC, Ctr Biol Mol Severo Ochoa, E-28049 Madrid, Spain
关键词
D O I
10.1074/jbc.M110294200
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
In this study we have used the yeast two-hybrid system to identify proteins that interact with the carboxyl-cytoplasmic domain (residues 464-509) of the insulin-sensitive glucose transporter GLUT4 (C-GLUT4). Using as bait C-GLUT4, we have isolated the carboxyl domain of Daxx (C-Daxx), the adaptor protein associated with the Fas and the type 11 TGF-beta (TbetaRII) receptors (1, 2). The two-hybrid interaction between C-GLUT4 and C-Daxx is validated by the ability of in vitro translated C-GLUT4 to interact with in vitro translated full-length Daxx and C-Daxx. C-Daxx does not interact with the C-cytoplasmic domain of GLUT1, the ubiquitous glucose transporter homologous to GLUT4. Replacement of alanine and serine for the dileucine pair (Leu(419)-Leu(490)) critical for targeting GLUT4 from the trans-Golgi network to the perinuclear intracellular store as well as for its surface internalization by endocytosis inhibits 2-fold the interaction of C-GLUT4 with Daxx. Daxx is pulled down with GLUT4 immunoprecipitated from lysates of 3T3-L1 fibroblasts stably transfected with GLUT4 and 3T3-L1 adipocytes expressing physiological levels of the two proteins. Similarly, GLUT4 is recovered with anti-Daxx immunoprecipitates. Using an established cell fractionation procedure we present evidence for the existence of two distinct intracellular Daxx pools in the nucleus and low density microsomes. Confocal immunofluorescence microscopy studies localize Daxx to promyelocytic leukemia nuclear bodies and punctate cytoplasmic structures, often organized in strings and underneath the plasma membrane. Daxx and GLUT4 are SUMOlated as shown by their reaction with an anti-SUM01 antibody and by the ability of this antibody to pull down Daxx and GLUT4.
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页码:19783 / 19791
页数:9
相关论文
共 36 条
[1]   Hepatitis delta virus replication generates complexes of large hepatitis delta antigen and antigenomic RNA that affiliate with and alter nuclear domain 10 [J].
Bell, P ;
Brazas, R ;
Ganem, D ;
Maul, GG .
JOURNAL OF VIROLOGY, 2000, 74 (11) :5329-5336
[2]  
BORDIER C, 1981, J BIOL CHEM, V256, P1604
[3]   Activation of apoptosis signal regulating kinase 1 (ASK1) by the adapter protein Daxx [J].
Chang, HY ;
Nishitoh, H ;
Yang, XL ;
Ichijo, H ;
Baltimore, D .
SCIENCE, 1998, 281 (5384) :1860-1863
[4]   A DOUBLE LEUCINE WITHIN THE GLUT4 GLUCOSE-TRANSPORTER COOH-TERMINAL DOMAIN FUNCTIONS AS AN ENDOCYTOSIS SIGNAL [J].
CORVERA, S ;
CHAWLA, A ;
CHAKRABARTI, R ;
JOLY, M ;
BUXTON, J ;
CZECH, MP .
JOURNAL OF CELL BIOLOGY, 1994, 126 (04) :979-989
[5]   EXOFACIAL EPITOPE-TAGGED GLUCOSE-TRANSPORTER CHIMERAS REVEAL COOH-TERMINAL SEQUENCES GOVERNING CELLULAR-LOCALIZATION [J].
CZECH, MP ;
CHAWLA, A ;
WOON, CW ;
BUXTON, J ;
ARMONI, M ;
TANG, W ;
JOLY, M ;
CORVERA, S .
JOURNAL OF CELL BIOLOGY, 1993, 123 (01) :127-135
[6]  
CZECH MP, 1993, J BIOL CHEM, V1, P127
[7]  
Everett RD, 1999, J CELL SCI, V112, P3443
[8]   Functional organization of the yeast proteome by systematic analysis of protein complexes [J].
Gavin, AC ;
Bösche, M ;
Krause, R ;
Grandi, P ;
Marzioch, M ;
Bauer, A ;
Schultz, J ;
Rick, JM ;
Michon, AM ;
Cruciat, CM ;
Remor, M ;
Höfert, C ;
Schelder, M ;
Brajenovic, M ;
Ruffner, H ;
Merino, A ;
Klein, K ;
Hudak, M ;
Dickson, D ;
Rudi, T ;
Gnau, V ;
Bauch, A ;
Bastuck, S ;
Huhse, B ;
Leutwein, C ;
Heurtier, MA ;
Copley, RR ;
Edelmann, A ;
Querfurth, E ;
Rybin, V ;
Drewes, G ;
Raida, M ;
Bouwmeester, T ;
Bork, P ;
Seraphin, B ;
Kuster, B ;
Neubauer, G ;
Superti-Furga, G .
NATURE, 2002, 415 (6868) :141-147
[9]   The sentrin-conjugating enzyme mUbc9 interacts with GLUT4 and GLUT1 glucose transporters and regulates transporter levels in skeletal muscle cells [J].
Giorgino, F ;
de Robertis, O ;
Laviola, L ;
Montrone, C ;
Perrini, S ;
McCowen, KC ;
Smith, RJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (03) :1125-1130