PubMed Hybridoma cell lines were used for the production of rat monoclonal immunoglobulin G (IgG) antibodies against MLV SU (83A25, kindly provided by L.H.Evans ). Here we provide evidence for the association of the envelope protein (Env) of the 4070A isolate of amphotropic murine leukaemia virus (A-MLV) with lipid rafts. We inserted all amino acid epitope of the VSV G protein at various locations including the extracellular loop 5 and the C-terminal extremity. Science 266, 1373–1376. 220.127.116.11. 10.1073/pnas.240459697, Lindwasser OW, Resh MD: Multimerization of human immunodeficiency virus type 1 Gag promotes its localization to barges, raft-like membrane microdomains. 10.1128/JVI.75.15.7175-7183.2001 PubMedCentralCrossRefPubMed, Zurück zum Zitat Sandrin V, Muriaux D, Darlix JL, Cosset FL: Intracellular trafficking of Gag and Env proteins and their interactions modulate pseudotyping of retroviruses. 10.1146/annurev.biochem.67.1.199 CrossRefPubMed, Zurück zum Zitat Schmid SL, Smythe E: Stage-specific assays for coated pit formation and coated vesicle budding in vitro. Cav-1 is also present in lipid rafts, which are thought to be precursors of caveolae ("pre-caveolae") . The temperature stability of mouse retroviruses depends on the cholesterol levels of viral lipid shell and cellular plasma membrane. J Virol 1990, 64: 6176-6183. J Virol 2003, 77: 1916-1926. Targeting retroviral vectors to specific cells. D) NIH3T3 cells (Env negative) stained for A-MLV Env, negative control (see text for details). A neutralizable epitope common to the envelope glycoproteins of ecotropic, polytropic, xenotropic, and amphotropic murine leukemia viruses. Wang, H., Paul, R., Burgeson, R E., Keene, D. R. and Kabat, D. (1991b). New York: Cold Spring Harbor Lab. Zurück zum Zitat Nichols BJ: GM1-containing lipid rafts are depleted within clathrin-coated pits. The specific lipid composition of DRMs leads to the selective incorporation and concentration of specific cellular proteins (reviewed in ). A-MLV Env was detected using antibodies produced by the hybridoma cell line 83A25. After washing once with PBS, the cells were overlaid with 200 μl PBS or 0.5% Triton X-100 (4°C) and incubated for 1 minute at 4°C (gently shaking at 8 rpm). J. Virol. 10.1016/0378-1119(88)90506-9 CrossRefPubMed. J Cell Biol 1991, 114: 869-880. J. Virol 28, 686–696. 10.1006/viro.1998.9398, Wirth M, Bode J, Zettlmeisl G, Hauser H: Isolation of overproducing recombinant mammalian cell lines by a fast and simple selection procedure. Receptor choice determinants in the envelope glycoproteins of amphotropic, xenotropic and polytropic murine leukemia viruses. 69, 6314–6322. The samples were overlaid with a discontinuous sucrose or OptiPrep gradient (35% – 5%). 10.1128/JVI.75.15.7175-7183.2001, Sandrin V, Muriaux D, Darlix JL, Cosset FL: Intracellular trafficking of Gag and Env proteins and their interactions modulate pseudotyping of retroviruses. Pinter, A., Chen, T. E., Lowy, A., Cortez, N. G. and Siligari, S. (1986). CB conceived of the study, carried out the experimental work and helped to draft the manuscript. The amino-terminal domain of murine leukemia virus (MuLV) envelope glycoproteins (SU) is sufficient for binding cell surface receptors and mediating entry into cells. Sequence archive. Virus entry was completely abolished in the presence of cytochalasine. Molecular Biotechnology GBF, German Research Centre for Biotechnology, Mascheroder Weg 1, Braunschweig, D, 38124, Germany, Institute of Clinical Medicine and Department of Molecular Biology, University of Aarhus, Aarhus, Denmark, You can also search for this author in Proc Natl Acad Sci USA 2000, 97: 13523-13525. The gradient was centrifuged at 4°C with 38000 rpm for 20 h. Six fractions were collected from the top of the tube. Sequences determining the pH dependence of viral entry are distinct from the host range-determining region of the murine ecotropic and amphotropic retrovirus envelope proteins. Zurück zum Zitat Nguyen DH, Hildreth JE: Evidence for budding of human immunodeficiency virus type 1 selectively from glycolipid-enriched membrane lipid rafts. Kabat, D. (1995). Identification of a subdomain in the Moloney murine leukemia virus envelope protein involved in receptor binding. Confirming the routine of the fractionation experiments, unmodified eGFP, which is localized in the cytoplasm, was exclusively found in the soluble fractions 5 and 6 (Fig 1A). Aloia RC, Tian H, Jensen FC: Lipid composition and fluidity of the human immunodeficiency virus envelope and host cell plasma membranes. GM1-containing lipid rafts are depleted within clathrin-coated pits. 28, 819–827. J. Virol. Article 10.1128/JVI.78.13.7153-7164.2004 PubMedCentralCrossRefPubMed. PubMedCentralCrossRefPubMed. 69, 3433–3440. J. Virol. 67, 4056–4061. PubMed Central With respect to the distribution in the plasma membrane, TX-100 resistance, and MBCD extraction, A-MLV Env exhibits similar properties as the DRM marker GM1 and distinct properties compared to CD71. Here we provide evidence for the association of the envelope protein (Env) of the 4070A isolate of amphotropic murine leukaemia virus (A-MLV) with lipid rafts. 66, 1468–1475. To investigate the association of proteins with cholesterol-rich microdomains via Western or Dot Blot the extraction of TX-100 soluble proteins was performed as described previously  with the following modifications. 66, 2281–2287. This is not uncommon for DRM-associated proteins as it has been shown for, e.g., HIV-1 p17 and gp41 . Mutational analysis of the envelope gene of Moloney murine leukemia virus. Our finding may have consequences for the understanding of A-MLV assembly and budding, which is known to be a specific and coordinated process. Zurück zum Zitat Lindwasser OW, Resh MD: Multimerization of human immunodeficiency virus type 1 Gag promotes its localization to barges, raft-like membrane microdomains. By using this website, you agree to our J Virol 2000, 74: 9911-9915. Six fractions were collected from the top of the tube. PubMed pMLVampho contains the complete genome of A-MLV cloned into pBluescript (Genethon, France received via J.-C. Proc Natl Acad Sci USA 1993, 90: 5181-5185. Intracellular trafficking of Gag and Env proteins and their interactions modulate pseudotyping of retroviruses.
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