Fluorescence Resonance Energy Transfer Imaging Reveals that Chemokine-Binding Modulates Heterodimers of CXCR4 and CCR5 Receptors

Isik, Nilgun; Hereld, Dale; Jin, Tian

doi:10.1371/journal.pone.0003424

Cited by 45 publications

(41 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This fact is the case of the two main HIV-1 coreceptors, CXCR4 and CCR5. When coexpressed on a cell and in the absence of ligands, these two receptors form heterodimers (39,40,44) that appear to modulate lymphocyte functions (40). This effect is compatible with the consensus for the G protein-coupled receptors (GPCR), which considers heteromers as entities whose function differs from that of the individual receptors (45).…”

Section: Ccr5 Expression In Cd4/cxcr4 Cells Blocks Gp120 Iiib -Inducementioning

confidence: 55%

CCR5/CD4/CXCR4 oligomerization prevents HIV-1 gp120_IIIBbinding to the cell surface

Martínez-Muñoz

Barroso

Dyrhaug

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

CCR5 and CXCR4, the respective cell surface coreceptors of R5 and X4 HIV-1 strains, both form heterodimers with CD4, the principal HIV-1 receptor. Using several resonance energy transfer techniques, we determined that CD4, CXCR4, and CCR5 formed heterotrimers, and that CCR5 coexpression altered the conformation of both CXCR4/CXCR4 homodimers and CD4/CXCR4 heterodimers. As a result, binding of the HIV-1 envelope protein gp120 IIIB to the CD4/CXCR4/CCR5 heterooligomer was negligible, and the gp120-induced cytoskeletal rearrangements necessary for HIV-1 entry were prevented. CCR5 reduced HIV-1 envelope-induced CD4/ CXCR4-mediated cell-cell fusion. In nucleofected Jurkat CD4 cells and primary human CD4 + T cells, CCR5 expression led to a reduction in X4 HIV-1 infectivity. These findings can help to understand why X4 HIV-1 strains infection affect T-cell types differently during AIDS development and indicate that receptor oligomerization might be a target for previously unidentified therapeutic approaches for AIDS intervention.chemokine receptors | oligomer formation | FRET/BRET F or HIV-1 to enter a target cell, the viral envelope glycoprotein gp120 must interact with a set of cell surface molecules that include the primary receptor, CD4 (1), and a chemokine receptor (CCR5 or CXCR4) that acts as a coreceptor (2, 3). These molecules form CD4/chemokine receptor complexes, as deduced from coprecipitation data for CXCR4 or CCR5 with CD4 (4-8).Most HIV-1 variants isolated from newly infected individuals use CCR5 and CD4 to enter host cells; these M-tropic R5 strains are predominant in acute and asymptomatic phases of HIV infection. CD4 + T helper type 1 (Th1) cells, which express high CCR5 levels (9, 10), are implicated in maintaining asymptomatic status (11, 12). The "viral shift" from R5 to T-tropic X4 HIV-1 strains correlates with AIDS progression (13,14). X4 strains infect mainly CD4 + Th2 cells, which express little CCR5 and whose CXCR4 levels resemble those of Th1 cells (15,16), which suggests that cell susceptibility to HIV-1 infection depends on the CD4/coreceptor ratio and on receptor levels during cell activation and/or differentiation (17). CXCR4 and CCR5 are present as homodimers and heterodimers at the plasma membrane (18)(19)(20). In addition, gp120-mediated CD4/CXCR4 and CD4/CCR5 association and clustering is reported (21-23). Nonetheless, little is known of how CCR5 expression influences the CD4/CXCR4 interaction, or of the molecular basis that underlies the differences in X4 strains infection relative to CCR5 levels at the cell surface.Here, we identify CD4/CXCR4/CCR5 oligomers at the cell membrane, even in the absence of ligands. CCR5 expression in these complexes modifies the heterodimeric CD4/CXCR4 conformation and blocks gp120 IIIB binding, without altering binding of the CXCR4 ligand CXCL12 and its subsequent signaling. gp120 IIIB -triggered LIMK1 activation, cofilin dephosphorylation, and the actin cytoskeleton rearrangement necessary for cell-cell fusion were impeded in CD4/CXCR4/CCR5-expressing c...

show abstract

Section: Ccr5 Expression In Cd4/cxcr4 Cells Blocks Gp120 Iiib -Inducementioning

confidence: 55%

CCR5/CD4/CXCR4 oligomerization prevents HIV-1 gp120_IIIBbinding to the cell surface

Martínez-Muñoz

Barroso

Dyrhaug

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…The finding that CCR5 antagonists block FBS-induced invasion is novel and suggested that CCR5 activation contribute to the production of metastasis in vivo where different chemotactic and growth signals are present. The mechanisms involved in CCR5 regulation of FBS-activated invasiveness are uncharacterized but they may include heterodimerization and ligand affinity regulation of other GPCRs (46), or the transactivation of growth factor receptor-(47) or integrin-mediated signaling (48), as described in noncancerous cells. The in vivo antimetastatic effect of maraviroc was shown by injecting MDA.pFULG cells into the circulation of immunodeficient mice and treating them with clinically relevant doses of the drug.…”

Section: Discussionmentioning

confidence: 99%

CCR5 Antagonist Blocks Metastasis of Basal Breast Cancer Cells

et al. 2012

View full text Add to dashboard Cite

The roles of the chemokine CCL5 and its receptor CCR5 in breast cancer progression remain unclear. Here, we conducted microarray analysis on 2,254 human breast cancer specimens and found increased expression of CCL5 and its receptor CCR5, but not CCR3, in the basal and HER-2 genetic subtypes. The subpopulation of human breast cancer cell lines found to express CCR5 displayed a functional response to CCL5. In addition, oncogene transformation induced CCR5 expression, and the subpopulation of cells that expressed functional CCR5 also displayed increased invasiveness. The CCR5 antagonists maraviroc or vicriviroc, developed to block CCR5 HIV coreceptor function, reduced in vitro invasion of basal breast cancer cells without affecting cell proliferation or viability, and maraviroc decreased pulmonary metastasis in a preclinical mouse model of breast cancer. Taken together, our findings provide evidence for the key role of CCL5/CCR5 in the invasiveness of basal breast cancer cells and suggest that CCR5 antagonists may be used as an adjuvant therapy to reduce the risk of metastasis in patients with the basal breast cancer subtype. Cancer Res; 72(15); 3839-50. Ó2012 AACR.

show abstract

“…Dimerization of a WT CXCR4 with a C-terminally truncated mutant causing the "warts, hypogammaglobulinemia, infections and myelokathexis" (WHIM) syndrome has been implicated in its resistance to desensitization and enhanced signaling in heterozygous WHIM patients (27). CXCR4 has also been shown to heterodimerize with other chemokine receptors and with GPCRs outside the chemokine family (28)(29)(30)(31)(32), with consequences including transinhibition of ligand binding (28) and changes in G protein and β-arrestin coupling…”

mentioning

confidence: 99%

Stoichiometry and geometry of the CXC chemokine receptor 4 complex with CXC ligand 12: Molecular modeling and experimental validation

Kufareva

Stephens

Holden

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

124

View full text Add to dashboard Cite

Chemokines and their receptors regulate cell migration during development, immune system function, and in inflammatory diseases, making them important therapeutic targets. Nevertheless, the structural basis of receptor:chemokine interaction is poorly understood. Adding to the complexity of the problem is the persistently dimeric behavior of receptors observed in cell-based studies, which in combination with structural and mutagenesis data, suggest several possibilities for receptor:chemokine complex stoichiometry. In this study, a combination of computational, functional, and biophysical approaches was used to elucidate the stoichiometry and geometry of the interaction between the CXC-type chemokine receptor 4 (CXCR4) and its ligand CXCL12. First, relevance and feasibility of a 2:1 stoichiometry hypothesis was probed using functional complementation experiments with multiple pairs of complementary nonfunctional CXCR4 mutants. Next, the importance of dimers of WT CXCR4 was explored using the strategy of dimer dilution, where WT receptor dimerization is disrupted by increasing expression of nonfunctional CXCR4 mutants. The results of these experiments were supportive of a 1:1 stoichiometry, although the latter could not simultaneously reconcile existing structural and mutagenesis data. To resolve the contradiction, cysteine trapping experiments were used to derive residue proximity constraints that enabled construction of a validated 1:1 receptor: chemokine model, consistent with the paradigmatic two-site hypothesis of receptor activation. The observation of a 1:1 stoichiometry is in line with accumulating evidence supporting monomers as minimal functional units of G protein-coupled receptors, and suggests transmission of conformational changes across the dimer interface as the most probable mechanism of altered signaling by receptor heterodimers.chemokine receptor | GPCR dimerization | molecular docking | functional complementation | cysteine trapping T he chemokine receptor CXCR4 regulates cell migration during many developmental processes (1, 2). Along with CCR5, it serves as one of the principal coreceptors for HIV entry into leukocytes (3), and is one of the most important chemokine receptors involved in cancer metastasis (4). Stromal-cell derived factor 1 (SDF-1 or CXCL12) was its only known ligand until recently, when CXCR4 was also shown to bind CXCL14 (5) and extracellular ubiquitin (6). Although structures of CXCR4 (7) and CCR5 (8) have been solved with synthetic antagonists, the structural basis for the interaction of CXCR4 (or any other chemokine receptor) with their natural ligands has yet to be determined. Numerous mutagenesis and NMR studies indicate that receptor:chemokine interactions involve two distinct sites (9-12), which has led to a two-site hypothesis of receptor activation (13). The so-called chemokine recognition site 1 (CRS1) (14) includes the N terminus of the receptor interacting with the globular core of the chemokine, whereas chemokine recognition site 2 (CRS2), located within the tra...

show abstract

Fluorescence Resonance Energy Transfer Imaging Reveals that Chemokine-Binding Modulates Heterodimers of CXCR4 and CCR5 Receptors

Cited by 45 publications

References 27 publications

CCR5/CD4/CXCR4 oligomerization prevents HIV-1 gp120_IIIBbinding to the cell surface

CCR5/CD4/CXCR4 oligomerization prevents HIV-1 gp120_IIIBbinding to the cell surface

CCR5 Antagonist Blocks Metastasis of Basal Breast Cancer Cells

Stoichiometry and geometry of the CXC chemokine receptor 4 complex with CXC ligand 12: Molecular modeling and experimental validation

Contact Info

Product

Resources

About

Fluorescence Resonance Energy Transfer Imaging Reveals that Chemokine-Binding Modulates Heterodimers of CXCR4 and CCR5 Receptors

Cited by 45 publications

References 27 publications

CCR5/CD4/CXCR4 oligomerization prevents HIV-1 gp120IIIBbinding to the cell surface

CCR5/CD4/CXCR4 oligomerization prevents HIV-1 gp120IIIBbinding to the cell surface

CCR5 Antagonist Blocks Metastasis of Basal Breast Cancer Cells

Stoichiometry and geometry of the CXC chemokine receptor 4 complex with CXC ligand 12: Molecular modeling and experimental validation

Contact Info

Product

Resources

About

CCR5/CD4/CXCR4 oligomerization prevents HIV-1 gp120_IIIBbinding to the cell surface

CCR5/CD4/CXCR4 oligomerization prevents HIV-1 gp120_IIIBbinding to the cell surface