Pleiotrophin inhibits HIV infection by binding the cell surface-expressed nucleolin

Université Paris-Est Créteil Val de Marne - Université Paris 12, Créteil, Île-de-France, France
FEBS Journal (Impact Factor: 4). 10/2005; 272(18):4646-59. DOI: 10.1111/j.1742-4658.2005.04870.x
Source: PubMed


The growth factor pleiotrophin (PTN) has been reported to bind heparan sulfate and nucleolin, two components of the cell surface implicated in the attachment of HIV-1 particles to cells. Here we show that PTN inhibits HIV-1 infection by its capacity to inhibit HIV-1 particle attachment to the surface of permissive cells. The beta-sheet domains of PTN appear to be implicated in this inhibitory effect on the HIV infection, in particular the domain containing amino acids 60-110. PTN binding to the cell surface is mediated by high and low affinity binding sites. Other inhibitors of HIV attachment known to bind specifically surface expressed nucleolin, such as the pseudopeptide HB-19 and the cytokine midkine prevent the binding of PTN to its low affinity-binding site. Confocal immunofluorescence laser microscopy revealed that the cross-linking of surface-bound PTN with a specific antibody results in the clustering of cell surface-expressed nucleolin and the colocalization of both PTN and nucleolin signals. Following its binding to surface-nucleolin, PTN is internalized by a temperature sensitive mechanism, a process which is inhibited by HB-19 and is independent of heparan and chondroitin sulfate proteoglycans. Nevertheless, proteoglycans might play a role in the concentration of PTN on the cell surface for a more efficient interaction with nucleolin. Our results demonstrate for the first time that PTN inhibits HIV infection and suggest that the cell surface-expressed nucleolin is a low affinity receptor for PTN binding to cells and it is also implicated in PTN entry into cells by an active process.

Download full-text


Available from: Elias Said, Oct 13, 2014
  • Source
    • "NCL controls a wide range of fundamental cellular processes such as ribosome biogenesis, proliferation and cellular cycle regulation (Cong et al., 2012; Ginisty et al., 1998; Ugrinova et al., 2007). On the cell surface, NCL serves as a receptor for ligands such as lactoferrin (Legrand et al., 2004), midkine protein (MK) (Take et al., 1994), pleiotrophin (PTN) (Said et al., 2005), and AGRO100, an oligonucleotide aptamer that has been shown to bind with high specificity and affinity to the NCL RRMs (Bates et al., 2009; Dahl et al., 1987; Mongelard and Bouvet, 2010; Reyes-Reyes et al., 2010; Soundararajan et al., 2009; Xu et al., 2001). The cell surfaceexpressed NCL is required for the efficient entry of human parainfluenza virus type 3 (HPV 3) into human lung epithelial A549 cells (Bose et al., 2004). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Feline calicivirus depends on host-cell proteins for its replication. We previously showed that knockdown of nucleolin (NCL), a phosphoprotein involved in ribosome biogenesis, resulted in the reduction of FCV protein synthesis and virus yield. Here, we found that NCL may not be involved in FCV binding and entry into cells, but it binds to both ends of the FCV genomic RNA, and stimulates its translation in vitro. AGRO100, an aptamer that specifically binds and inactivates NCL, caused a strong reduction in FCV protein synthesis. This effect could be reversed by the addition of full-length NCL but not by a ΔrNCL, lacking the N-terminal domain. Consistent with this, FCV infection of CrFK cells stably expressing ΔrNCL led to a reduction in virus protein translation. These results suggest that NCL is part of the FCV RNA translational complex, and that the N-terminal part of the protein is required for efficient FCV replication.
    Full-text · Article · Feb 2016 · Virology
  • Source
    • "Recognition of early apoptotic cells by cell surface NCL on monocytes and macrophages [139] [140] MK Temperature-dependent active internalization process / Inhibition of HIV-1 infection [42] [43] [142] HGF Regulatory interplay between stromal and epithelial cells within the prostate [102] VEGF VEGF 165 -induced endothelial cell migration [106] [122] PTN Temperature-dependent active internalization process / Inhibition of HIV-1 infection / PTNinduced endothelial cell migration [143] [116] Acharan sulfate Anti-tumor activity in vitro and in vivo [101] ADAMTS-2 Anti-angiogenic action in vitro and in vivo [147] Lactoferrin Inhibition of the proliferation of cancerous mammary gland epithelial cells / Potent antiviral activity against HIV and CMV [44] ES Transportation of ES in the nucleus / Antiangiogenic and anti-tumor activities of ES in vivo / Anti-lymphangiogenic activities of ES [98, 148] scuPA Transportation of scuPA in the nucleus / Induction of α-SMA expression in human fibroblasts [40] Tumor-homing peptide F3 "
    [Show abstract] [Hide abstract]
    ABSTRACT: A large number of mostly recent reports show enhanced expression of the multi-functional protein nucleolin (NCL) on the surface of activated lymphocytes, angiogenic endothelial and many different types of cancer cells. Translocation of NCL at the external side of the plasma membrane occurs via a secretory pathway independent of the endoplasmic reticulum-Golgi complex, requires intracellular intact actin cytoskeleton, and seems to be mediated by a variety of factors. Cell surface NCL serves as a binding partner of several molecules implicated in cell differentiation, adhesion, leukocyte trafficking, inflammation, angiogenesis and tumor development, mediating their biological activities and in some cases, leading to their internalization. Accumulating evidence validates cell surface NCL as a strategic target for treatment of cancer, while its property of tumor-specific uptake of targeted ligands seems to be useful for the development of non-invasive imaging tools for the diagnosis of cancer and for the targeted release of chemotherapeutic drugs. The observation that cell surface NCL exists in complexes with several other proteins implicated in tumorigenesis and angiogenesis suggests that targeting cell surface NCL might trigger multi-inhibitory effects, depending on the cell type. This review summarizes papers and patents related to the redistribution and the biological functions of cell surface NCL, with emphasis on the potential importance and advantages of developing efficient anti-cell surface NCL strategies.
    Full-text · Article · Nov 2013
  • Source
    • "Nuclear translocation of PTN has been previously discussed, based on the observations that its primary structure contains three potential nuclear targeting sequences [32], it was found to bind to NCL [33], and NCL was found to participate in midkine nuclear translocation [10,33]. Although interaction of PTN with NCL has been previously mentioned [8], it is the first time that it is clearly shown that NCL participates in PTN nuclear translocation. At the moment, the functional significance of the nuclear localization of PTN is completely unknown. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Pleiotrophin (PTN) is a heparin-binding growth factor with significant role(s) in tumour growth and angiogenesis. Although implication of endogenous PTN has been studied in several in vivo models of tumour angiogenesis, its role in physiological angiogenesis has not been addressed. In the present work, we studied expression and functional significance of endogenous PTN during angiogenesis in the chicken embryo chorioallantoic membrane (CAM). Using molecular, cellular and biochemical assays, we studied the expression pattern of PTN in CAM and human endothelial cells and its possible interaction with nucleolin (NCL). CAM cells were transfected with a pCDNA3.1 vector, empty (PC) or containing full length cDNA for PTN in antisense orientation (AS-PTN). Angiogenesis was estimated by measuring total vessel length. In vitro, human endothelial cells migration was studied by using a transwell assay, and down-regulation of NCL was performed by using a proper siRNA. Endogenous PTN mRNA and protein levels, as well as protein levels of its receptor protein tyrosine phosphatase beta/zeta (RPTPβ/ζ) were maximal at early stages, when CAM angiogenesis is active. Application of AS-PTN onto CAM at days of active angiogenesis was not toxic to the tissue and led to dose-dependent decreased expression of endogenous PTN, ERK1/2 activity and angiogenesis. Interestingly, endogenous PTN was also immunolocalized at the endothelial cell nucleus, possibly through interaction with NCL, a protein that has a significant role in the nuclear translocation of many proteins. Down-regulation of NCL by siRNA in human endothelial cells significantly decreased nuclear PTN, verifying this hypothesis. Moreover, it led to abolishment of PTN-induced endothelial cell migration, suggesting, for the first time, that PTN-NCL interaction has a functional significance. Expression of endogenous PTN correlates with and seems to be involved in angiogenesis of the chicken embryo CAM. Our data suggest that NCL may have a role, increasing the number of growth factors whose angiogenic/tumorigenic activities are mediated by NCL.
    Full-text · Article · Mar 2012 · Vascular Cell
Show more