![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
CELLULAR AND MOLECULAR
and the HIV-1 Coat Protein gp120: A Fluorescence Resonance Energy Transfer (FRET) Study
Department of Molecular Pharmacology and Biological Chemistry, The Feinberg School of Medicine, Northwestern University, Chicago, Illinois
Both the chemokine SDF-1
and the human immunodeficiency virus-1 (HIV-1) coat protein gp120 can bind to CXCR4 chemokine receptors but with different signaling consequences. To understand the molecular basis for these differences, we tagged the rat CXCR4 receptor with enhanced cyan (ECFP) and yellow (EYFP) derivatives of the green fluorescent protein and investigated CXCR4 receptor dimerization in human embryonic kidney (HEK)-tsA201 cells using fluorescence resonance energy transfer (FRET). Elevated FRET was detected under basal conditions from EYFP-CXCR4 and ECFP-CXCR4 receptor-transfected cells indicating a high level of CXCR4 receptor dimerization. In comparison, EYFP-CXCR4 and ECFP-µ-opioid receptor-cotransfected cells displayed a much lower FRET signal. The FRET signal resulting from EYFP-CXCR4- and ECFP-CXCR4-expressing cells could be attenuated by coexpressing nontagged CXCR4 receptors suggesting competition with fluorophore-tagged receptors in the membrane. Nontagged µ-opioid, 
-opioid, and muscarinic receptors also decreased the FRET between the tagged CXCR4 receptor pairs but to a lesser extent. Application of the CXCR4 receptor agonist SDF-1 (50 nM) further increased the FRET signal from tagged CXCR4 receptors, an effect that was inhibited by the CXCR4 antagonist AMD3100. SDF-1 had no effect when EYFP-CXCR4 and ECFP-µ-opioid receptors were coexpressed. The effect of gp120IIIB on CXCR4 FRET was dependent on the coexpression of human CD4 (hCD4) when it increased the FRET signal, and this was decreased by AMD3100 pretreatment. FRET analysis of tagged hCD4 constructs demonstrated that there was significant association of hCD4 and CXCR4, as well as hCD4 dimerization. These data suggest that CXCR4 dimerization is involved in SDF-1- and gp120-induced signaling events.
Address correspondence to: Dr. Richard J. Miller, Department of Molecular Pharmacology and Biological Chemistry, Northwestern University, 303 E. Chicago Ave, Chicago, IL 60611. E-mail: r-miller10{at}northwestern.edu
This article has been cited by other articles:
|
|
 |
|
  L. Navarro-Borelly, A. Somasundaram, M. Yamashita, D. Ren, R. J. Miller, and M. Prakriya STIM1-Orai1 interactions and Orai1 conformational changes revealed by live-cell FRET microscopy J. Physiol., November 15, 2008; 586(22): 5383 - 5401. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. J. Harris, M. J. Farquhar, C. J. Mee, C. Davis, G. M. Reynolds, A. Jennings, K. Hu, F. Yuan, H. Deng, S. G. Hubscher, et al. CD81 and Claudin 1 Coreceptor Association: Role in Hepatitis C Virus Entry J. Virol., May 15, 2008; 82(10): 5007 - 5020. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. A. White, H. Jung, and R. J. Miller Chemokines and the pathophysiology of neuropathic pain PNAS, December 18, 2007; 104(51): 20151 - 20158. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Chun and G. V. W. Johnson Activation of Glycogen Synthase Kinase 3beta Promotes the Intermolecular Association of Tau: THE USE OF FLUORESCENCE RESONANCE ENERGY TRANSFER MICROSCOPY J. Biol. Chem., August 10, 2007; 282(32): 23410 - 23417. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Guyon and J.-L. Nahon Multiple actions of the chemokine stromal cell-derived factor-1{alpha} on neuronal activity J. Mol. Endocrinol., March 1, 2007; 38(3): 365 - 376. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Gaibelet, T. Planchenault, S. Mazeres, F. Dumas, F. Arenzana-Seisdedos, A. Lopez, B. Lagane, and F. Bachelerie CD4 and CCR5 Constitutively Interact at the Plasma Membrane of Living Cells: A CONFOCAL FLUORESCENCE RESONANCE ENERGY TRANSFER-BASED APPROACH J. Biol. Chem., December 8, 2006; 281(49): 37921 - 37929. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Wang, L. He, C. A. Combs, G. Roderiquez, and M. A. Norcross Dimerization of CXCR4 in living malignant cells: control of cell migration by a synthetic peptide that reduces homologous CXCR4 interactions. Mol. Cancer Ther., October 1, 2006; 5(10): 2474 - 2483. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Zaitseva, T. Romantseva, J. Manischewitz, J. Wang, D. Goucher, and H. Golding Increased CXCR4-dependent HIV-1 fusion in activated T cells: role of CD4/CXCR4 association J. Leukoc. Biol., December 1, 2005; 78(6): 1306 - 1317. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Badr, G. Borhis, D. Treton, C. Moog, O. Garraud, and Y. Richard HIV Type 1 Glycoprotein 120 Inhibits Human B Cell Chemotaxis to CXC Chemokine Ligand (CXCL) 12, CC Chemokine Ligand (CCL)20, and CCL21 J. Immunol., July 1, 2005; 175(1): 302 - 310. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Percherancier, Y. A. Berchiche, I. Slight, R. Volkmer-Engert, H. Tamamura, N. Fujii, M. Bouvier, and N. Heveker Bioluminescence Resonance Energy Transfer Reveals Ligand-induced Conformational Changes in CXCR4 Homo- and Heterodimers J. Biol. Chem., March 18, 2005; 280(11): 9895 - 9903. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Ahr, M. Denizot, V. Robert-Hebmann, A. Brelot, and M. Biard-Piechaczyk Identification of the Cytoplasmic Domains of CXCR4 Involved in Jak2 and STAT3 Phosphorylation J. Biol. Chem., February 25, 2005; 280(8): 6692 - 6700. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M.-B. Huang, L. L. Jin, C. O. James, M. Khan, M. D. Powell, and V. C. Bond Characterization of Nef-CXCR4 Interactions Important for Apoptosis Induction J. Virol., October 15, 2004; 78(20): 11084 - 11096. [Abstract] [Full Text] [PDF]
|
|
 |
 |
 |
|
 |
 |
 |
