Document 0086
 DOCN  M9590086
 TI    In vitro binding and phosphorylation of human immunodeficiency virus
       type 1 Nef protein by serine/threonine protein kinase.
 DT    9509
 AU    Bodeus M; Marie-Cardine A; Bougeret C; Ramos-Morales F; Benarous R;
       INSERM U332, Institut Cochin de Genetique Moleculaire,; Universite Paris
       V-Rene Descartes, France.
 SO    J Gen Virol. 1995 Jun;76 ( Pt 6):1337-44. Unique Identifier : AIDSLINE
       MED/95302029
 AB    Although the human immunodeficiency virus type 1 (HIV-1) nef gene still
       has no precisely defined function, in vivo studies have demonstrated
       that Nef is an important pathogenic determinant of HIV. In order to
       identify cellular proteins capable of binding to Nef, the HIV-1LAI nef
       gene product was expressed in the bacterial vector pGEX-2T as a
       glutathione S-transferase (GST)-Nef fusion protein. Deletion mutants
       corresponding to 86 and 35 N-terminal residues of the Nef protein were
       prepared. The GST-Nef constructs were used to identify cellular kinases
       capable of interacting with Nef. After incubation with a Jurkat cell
       lysate, the GST-Nef constructs immobilized on glutathione-agarose beads
       bound to cellular kinase(s) and were phosphorylated at three sites in
       vitro: one on threonine at position 15, one on serine between residues 1
       and 35, and one on threonine between residues 36 and 86. The
       Nef-phosphorylating activity was inhibited by protein kinase C
       (PKC)-selective inhibitors. Cell fractionation showed that this
       Nef-binding kinase was mainly in the membrane-associated fraction. These
       results suggest that kinase(s) of the PKC family are specifically bound
       to and phosphorylate Nef in vitro. The interaction of Nef with cellular
       kinases and its phosphorylation may be important in mediating the
       effects of Nef in HIV-1 pathogenesis.
 DE    Base Sequence  Binding Sites  Cell Line  DNA Primers  Gene Products,
       nef/BIOSYNTHESIS/GENETICS/*METABOLISM  Genes, nef  Glutathione
       Transferases/BIOSYNTHESIS/METABOLISM  Human  HIV-1/GENETICS/*METABOLISM
       Kinetics  Molecular Sequence Data  Phosphorylation
       Phosphoserine/ANALYSIS  Phosphothreonine/ANALYSIS  Protein Binding
       Protein-Serine-Threonine Kinases/*METABOLISM  Recombinant Fusion
       Proteins/BIOSYNTHESIS/METABOLISM  Sequence Deletion  Support, Non-U.S.
       Gov't  Tyrosine/ANALOGS & DERIVATIVES/ANALYSIS  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).