The basic helix-loop-helix domain of the aryl hydrocarbon receptor nuclear transporter (ARNT) can oligomerize and bind E-box DNA specifically

被引:30
作者
Huffman, JL
Mokashi, A
Bächinger, HP
Brennan, RG
机构
[1] Oregon Hlth Sci Univ, Dept Biochem & Mol Biol, Portland, OR 97201 USA
[2] Portland Shriners Hosp, Portland, OR 97201 USA
关键词
D O I
10.1074/jbc.M105675200
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
The aryl hydrocarbon receptor nuclear transporter (ARNT) is a basic helix-loop-helix (bHLH) protein that contains a Per-Arnt-Sim (PAS) domain. ARNT heterodimerizes in vivo with other bHLH PAS proteins to regulate a number of cellular activities, but a physiological role for ARNT homodimers has not yet been established. Moreover, no rigorous studies have been done to characterize the biochemical properties of the bHLH domain of ARNT that would address this issue. To begin this characterization, we chemically synthesized a 56-residue peptide encompassing the bHLH domain of ARNT (residues 90-145). In the absence of DNA, the ARNT-bHLH peptide can form homodimers in lower ionic strength, as evidenced by dynamic light scattering analysis, and can bind E-box DNA (CACGTG) with high specificity and affinity, as determined by fluorescence anisotropy. Dimers and tetramers of ARNT-bHLH are observed bound to DNA in equilibrium sedimentation and dynamic light scattering experiments. The homodimeric peptide also undergoes a coil-to-helix transition upon E-box DNA binding. Peptide oligomerization and DNA affinity are strongly influenced by ionic strength. These biochemical and biophysical studies on the ARNT-bHLH reveal its inherent ability to form homodimers at concentrations supporting a physiological function and underscore the significant biochemical differences among the bHLH superfamily.
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页码:40537 / 40544
页数:8
相关论文
共 44 条
[1]   MOLECULAR CHARACTERIZATION OF HELIX-LOOP-HELIX PEPTIDES [J].
ANTHONYCAHILL, SJ ;
BENFIELD, PA ;
FAIRMAN, R ;
WASSERMAN, ZR ;
BRENNER, SL ;
STAFFORD, WF ;
ALTENBACH, C ;
HUBBELL, WL ;
DEGRADO, WF .
SCIENCE, 1992, 255 (5047) :979-983
[2]   CONSTITUTIVE FUNCTION OF THE BASIC HELIX-LOOP-HELIX PAS FACTOR ARNT - REGULATION OF TARGET PROMOTERS VIA THE E-BOX MOTIF [J].
ANTONSSON, C ;
ARULAMPALAM, V ;
WHITELAW, ML ;
PETTERSSON, S ;
POELLINGER, L .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (23) :13968-13972
[3]  
BASCI SG, 1996, J BIOL CHEM, V271, P8843
[4]  
BASCI SG, 1995, MOL PHARM, V47, P432
[5]   BASIC-HELIX-LOOP-HELIX REGION OF TAL - EVALUATION OF STRUCTURE AND DNA AFFINITY [J].
BISHOP, P ;
GHOSH, I ;
JONES, C ;
CHMIELEWSKI, J .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1995, 117 (31) :8283-8284
[6]   Expression of the gene encoding the proapoptotic Nip3 protein is induced by hypoxia [J].
Bruick, RK .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (16) :9082-9087
[7]   Mechanism of stabilization of helical conformations of polypeptides by water containing trifluoroethanol [J].
CammersGoodwin, A ;
Allen, TJ ;
Oslick, SL ;
McClure, KF ;
Lee, JH ;
Kemp, DS .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1996, 118 (13) :3082-3090
[8]   Role of HIF-1α or in hypoxia-mediated apoptosis, cell proliferation and tumour angiogenesis [J].
Carmeliet, P ;
Dor, Y ;
Herbert, JM ;
Fukumura, D ;
Brusselmans, K ;
Dewerchin, M ;
Neeman, M ;
Bono, F ;
Abramovitch, R ;
Maxwell, P ;
Koch, CJ ;
Ratcliffe, P ;
Moons, L ;
Jain, RK ;
Collen, D ;
Keshet, E .
NATURE, 1998, 394 (6692) :485-490
[9]  
COMPTON LA, 1987, J BIOL CHEM, V262, P13039
[10]   Control of cell lineage-specific development and transcription by bHLH-PAS proteins [J].
Crews, ST .
GENES & DEVELOPMENT, 1998, 12 (05) :607-620