A new HTLV-III/LAV Encoded Antigen Detected by Antibodies from AIDS Patients

Allan, Jonathan S.; Coligan, John E.; Lee, Tun‐Hou; McLane, MF; Kanki, Phyllis J.; Groopman, Jerome E.; Essex, M.

doi:10.1126/science.2997921

Cited by 212 publications

(110 citation statements)

References 31 publications

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“…The HIV-1 nef gene encodes a protein with an apparent molecular mass of 24-27 kDa, which is myristoylated (Allan et al, 1985;Guy et al, 1987). The biological function of Nef is not yet clear, however, it can be regarded as conclusive that it is not a guanine-nucleotide-binding protein (Kaminchik et al, 1990;Nebreda et al, 1991Nebreda et al, , 1992Matsuura et al, 1991;Backer et al, 1991;Harris et al, 1992;Wolber et al, 1992) participating in cellular signaling pathways as has been claimed by some authors (Samuel et al, 1987;Guy et al, 1987;Cheng-Mayer et al, 1989).…”

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confidence: 98%

Oligomerization of the Nef protein from human immunodeficiency virus (HIV) type 1

Kienzle

Freund

Kalbitzer

et al. 1993

European Journal of Biochemistry

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The nef genes, derived from two different human immunodeficiency-virus-type-1 (HIV-1) strains, were expressed in procaryotic cells (Escherichia coli) and in eucaryotic cells (insect cells infected with nef-containing baculovirus). The oligomerization of recombinant Nef protein was studied by NMR spectroscopy and immunoblotting under various experimental conditions. 1H-NMR spectroscopy shows that native folded protein has the tendency to polymerize under low-salt conditions. These oligomers become covalently linked by disulfide bonds after decreasing the reduction potential, a process which is fully reversible. Cross-linking studies with bis(sulfo-succinimidyl)suberate and alkylation with iodoacetic acid under non-reducing and reducing conditions document for the first time that Nef can also form homomeric structures including monomers, dimers, trimers and tetramers in cell lysates and intact cells. We found disulfide-linked as well as non-covalently associated oligomers. Since the Nef molecules are not exclusively found in the cytoplasm of HIV infected cells and since the reduced glutathione concentration in lymphocytes of virus infected persons is known to be unusually low, it might be possible that these Nef oligomers have a biological function in vivo as well

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confidence: 98%

Oligomerization of the Nef protein from human immunodeficiency virus (HIV) type 1

Kienzle

Freund

Kalbitzer

et al. 1993

European Journal of Biochemistry

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“…The nefgene product is myristylated at the N terminus (Allan et al, 1985) and this acylation is required for the location of nef on the cytoplasmic face of the plasma membrane in infected cells (Yu & Felsted, 1992). Myristylation of nef enhances HI¥-I replication (Zazapoulos & Haseltine, 1992) and, conversely, suppresses HIV-1 LTR transcription (Yu & Felsted, 1992).…”

Section: Introductionmentioning

confidence: 99%

“…Luria et al (1991) examined the effect of nefon the induction of interleukin 2 receptor (1R2R) mRNA: Jurkat T cells constitutively expressing nefwere unable to respond to a wide range of stimuli that normally induced the synthesis of IL2R mRNA. In contrast, cells expressing an alternative nef allele that differed at only three amino acid residues responded normally.The nefgene product is myristylated at the N terminus (Allan et al, 1985) and this acylation is required for the location of nef on the cytoplasmic face of the plasma membrane in infected cells (Yu & Felsted, 1992). Myristylation of nef enhances HI¥-I replication (Zazapoulos & Haseltine, 1992) and, conversely, suppresses HIV-1 LTR transcription (Yu & Felsted, 1992).…”

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Identification of cellular proteins that bind to the human immunodeficiency virus type 1 nef gene product in vitro: a role for myristylation

Harris

Coates

1993

Journal of General Virology

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The human immunodeficiency virus (HIV) type 1 nef gene product was expressed as an N-terminal fusion protein with glutathione-S-transferase (GST) in the baculovirus system. The resulting nefGST fusion protein was found to be authentically myristylated at the N terminus and could be purified to homogeneity by one-step affinity chromatography on immobilized glutathione. The high affinity of nefGST for glutathione was exploited to develop an assay to identify cellular proteins capable of interacting with nef Several such proteins were identified in extracts from the Jurkat human T cell line. The interaction between neJZbinding proteins and immobilized nefGST could be specifically competed by the addition of soluble nef Cell fractionation showed that neJ:binding proteins were present in both cytosolic and membrane-associated fractions. A non-myristylated derivative failed to bind to the membrane-associated proteins but was able to bind to the cytosolic group, albeit with reduced affinity. In addition, a single protein present in both soluble and membrane-associated fractions exhibited myristylation-independent binding to nef By analogy with other myristylated proteins such as MARCKS (myristylated alanine-rich C kinase substrate) and the Rous sarcoma virus transforming protein, src, the membrane-associated proteins that bind only to myristylated nef may represent a specific membrane target for nef The cytosolic proteins that interact with nef may constitute soluble components of an as yet unidentified signal transduction pathway which is the target of nefaction in the HIV-l-infected cell. IntroductionThe nef gene of human immunodeficiency virus (HIV) type 1 encodes a polypeptide of between 200 and 205 amino acids, varying between isolates (Harris et al., 1992a). Despite intensive work, the precise function of the nefgene product remains the subject of controversy. Initial studies identified the protein as a repressor of viral replication (Fisher et al., 1986; Luciw et al., 1987;Terwilliger et al., 1986) that acted by down-regulation of transcription from the HIV-1 long terminal repeat (LTR) (Ahmad & Venkatesan, 1988; Niederman et al., 1989;Maitra et al., 1991), but other studies failed to reproduce these results (Kim et al., 1989;Hammes et al., 1989; Bachelerie et al., 1990). There is a significant degree of amino acid sequence variation between nefisolates both in vivo and in vitro (Harris et al., 1992a) and the contradictions regarding nef function may represent the consequences of this variation. This problem was highlighted by a number of recent studies. Terwilliger et al. (1991) demonstrated that the replication of the IIIB provirus was suppressed by the corresponding IIIB nef allele but that replacement of nefin the IIIB provirus by the ELI nefallele enhanced the growth of the provirus. A number of amino acid residues in the ELI nef gene were subsequently identified as important for this enhancement, including Cys143, Cysl70 and Leu182 (Zazapoulos & Haseltine, 1992). Luria et al. (1991) examined the effect...

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“…Nef may be responsible for HIV evading the immune response [3]. In eucaryotic cells, Nef is usually myristoylated at its N-terminal glycine [5] and mainly localized in the cytoplasm associated with lipid membranes [6-91. However, Nef can also be found in the nucleus . It can be regarded as proven that Nef is not a guanine-nucleotide-binding protein .…”

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confidence: 99%

A possible regulation of negative factor (Nef) activity of human immunodeficiency virus type 1 by the viral protease

Freund

Kellner

Konvalinka

et al. 1994

European Journal of Biochemistry

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Negative factor (Nef) protein from human immunodeficiency virus type 1 (HIV-1) is cleaved into two well-defined domains by the HIV-1-encoded protease. The cleavage site is located between Trp57 and Leu58 and is well conserved. The two domains are stable in the presence of protease for more than 48 h. The C-terminal core domain contains a well-conserved well-folded region. The cleavage releases the core domain from the myristoylated membrane anchor domain. As is the case for other HIV proteins, cleavage of Nef could be crucial for correct biological function

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A new HTLV-III/LAV Encoded Antigen Detected by Antibodies from AIDS Patients

Cited by 212 publications

References 31 publications

Oligomerization of the Nef protein from human immunodeficiency virus (HIV) type 1

Oligomerization of the Nef protein from human immunodeficiency virus (HIV) type 1

Identification of cellular proteins that bind to the human immunodeficiency virus type 1 nef gene product in vitro: a role for myristylation

A possible regulation of negative factor (Nef) activity of human immunodeficiency virus type 1 by the viral protease

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