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    Cellular localization of Nef expressed in persistently HIV-1-infected low-producer astrocytes.
        
        Aids 6, 1427-1436 (1992)
    
    
    
				Objectives: The characterization and localization of HIV-1 Nef highly expressed in permanently infected astrocytes (TH4-7-5) as a model for latent infection of human brain cells. Design: Immunochemical methods are an appropriate tool to investigate expression and localization of cellular proteins. Methods: Nef expression was analysed by Western blot and immunoperoxidase staining using a panel of monoclonal and polyclonal antibodies. Cellular localization studies were performed by indirect immunofluorescence and subcellular fractionation of TH4-7-5 cells. Myristoylation of Nef was investigated by immunoprecipitation of [3H]myristic acid-labelled cell extract. TH4-7-5 nef gene was cloned and amplified by polymerase chain reaction and the nef nucleotide sequence analysed. Results: Reactivities of various Nef-specific antibodies with Nef antigen in TH4-7-5 cells were demonstrated by Western blot analysis. Immunofluorescence revealed cytoplasmic perinuclear staining of Nef with most antibodies. However, one monoclonal antibody against amino acids 168-175 of Nef showed intense homogeneous nuclear staining in TH4-7-5 cells. Reactivity of this Nef antibody was blocked with recombinant Nef derived from TH4-7-5 cells. After subcellular fractionation, Nef was detected in nuclear, membrane and cytosolic fractions of TH4-7-5 cells. No myristoylated Nef antigen was detectable, perhaps because of a serine residue at position 2 of the TH4-7-5 nef gene instead of the glycine residue required for myristoylation. Conclusions: Chronically HIV-1-infected astrocytoma cells with restricted virus production express different antigenic forms of Nef, which can be distinguished by their subcellular localization. Variant subcellular targeting of Nef suggests the existence of multiple activities of Nef within HIV-infected cells.
			
			
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        Publikationstyp
        Artikel: Journalartikel
    
 
    
        Dokumenttyp
        Wissenschaftlicher Artikel
    
 
     
    
    
        Schlagwörter
        Cellular Localization ; Hiv-1 Nef ; Human Astrocytoma Cells ; Myristoylation ; Nef Sequence ; Permanent Infection
    
 
     
    
    
        Sprache
        englisch
    
 
    
        Veröffentlichungsjahr
        1992
    
 
     
    
        HGF-Berichtsjahr
        0
    
 
    
    
        ISSN (print) / ISBN
        0269-9370
    
 
    
        e-ISSN
        1473-5571
    
 
     
     
     
	     
	 
	 
    
        Zeitschrift
        AIDS
    
 
		
    
        Quellenangaben
        
	    Band: 6,  
	    Heft: 12,  
	    Seiten: 1427-1436 
	    
	    
	
    
 
  
         
        
            Verlag
            Lippincott Williams & Wilkins
        
 
         
	
         
         
         
         
         
	
         
         
         
    
         
         
         
         
         
         
         
    
        Begutachtungsstatus
        Peer reviewed
    
 
    
        Institut(e)
        Institute of Virology (VIRO)
    
 
    
        POF Topic(s)
        30203 - Molecular Targets and Therapies
    
 
    
        Forschungsfeld(er)
        Immune Response and Infection
    
 
    
        PSP-Element(e)
        G-502700-001
    
 
     
     	
    
        Scopus ID
        0027082508
    
    
        Erfassungsdatum
        1992-12-31