Plasma membrane-targeted Raf kinase activates NF-κB and human immunodeficiency virus type 1 replication in T lymphocytes

被引:48
作者
Flory, E
Weber, CK
Chen, PF
Hoffmeyer, A
Jassoy, C
Rapp, UR
机构
[1] Univ Wurzburg, Inst Med Strahlenkunde & Zellforsch, D-97078 Wurzburg, Germany
[2] Univ Wurzburg, Inst Virol & Immunbiol, D-97078 Wurzburg, Germany
关键词
D O I
10.1128/JVI.72.4.2788-2794.1998
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Increasing evidence points to a role of the mitogenic Ras/Raf/MEK/ERK signaling cascade in regulation of human immunodeficiency virus type 1 (HIV-1) gene expression. Stimulation of elements of this pathway leads to transactivation of the HIV-1 promoter. In particular, the NF-kappa B motif in the HIV long terminal repeat (LTR) represents a Raf-responsive element in fibroblasts. Regulation of the Raf kinase in T cells differs from findings with a variety of cell lines that the catalytic domain of Raf (Raf(Delta 26-303)) shows no activity, In this study, we restored the activity of the kinase in T cells by fusing its catalytic domain to the CAAX motif (-Cx) of Ras,thus targeting the enzyme to the plasma membrane, Constitutive activity of Raf was demonstrated by phosphorylation of mitogen-activated protein kinase kinase (MEK) and endogenous mitogen-activated protein kinase 1/2 (ERK1/2) in A3.01 T cells transfected with Raf(Delta 26-303)-Cx. Membrane-targeted Raf also stimulates NF-kappa B, as judged by kappa B-dependent reporter assays and enhanced NF-kappa B p65 binding on band shift analysis. Moreover, we found that active Raf transactivates the HIVNL4-3 LTR in A3.01 T lymphocytes and that dominant negative Raf (C4) blocked 12-O-tetradecanoylphorbol-13-acetate induced transactivation. When cotransfected with infectious HIVNL4-3 DNA, membrane-targeted Raf induces viral replication up to 10-fold over basal levels, as determined by the release of newly synthesized p24(gag) protein, Our study clearly demonstrates that the activity of the catalytic domain of Raf in A3.01 T cells is dependent on its cellular localization, The functional consequences of active Raf in T lymphocytes include not only NF-kappa B activation and transactivation of the HIVNL4-3 LTR but also synthesis and release of HIV particles.
引用
收藏
页码:2788 / 2794
页数:7
相关论文
共 44 条
  • [11] Host factors and the pathogenesis of HIV-induced disease
    Fauci, AS
    [J]. NATURE, 1996, 384 (6609) : 529 - 534
  • [12] FINCO TS, 1993, J BIOL CHEM, V268, P17676
  • [13] Raf-1 kinase targets GA-binding protein in transcriptional regulation of the human immunodeficiency virus type 1 promoter
    Flory, E
    Hoffmeyer, A
    Smola, U
    Rapp, UR
    Bruder, JT
    [J]. JOURNAL OF VIROLOGY, 1996, 70 (04) : 2260 - 2268
  • [14] The Ras-Raf pathway is activated in human immunodeficiency virus-infected monocytes and participates in the activation of NF-kappa B
    Folgueira, L
    Algeciras, A
    MacMorran, WS
    Bren, GD
    Paya, CV
    [J]. JOURNAL OF VIROLOGY, 1996, 70 (04) : 2332 - 2338
  • [15] CELLULAR TRANSCRIPTION FACTORS INVOLVED IN THE REGULATION OF HIV-1 GENE-EXPRESSION
    GAYNOR, R
    [J]. AIDS, 1992, 6 (04) : 347 - 363
  • [16] GRILLI M, 1993, INT REV CYTOL, V143, P1
  • [17] MUTATIONAL ACTIVATION OF C-RAF-1 AND DEFINITION OF THE MINIMAL TRANSFORMING SEQUENCE
    HEIDECKER, G
    HULEIHEL, M
    CLEVELAND, JL
    KOLCH, W
    BECK, TW
    LLOYD, P
    PAWSON, T
    RAPP, UR
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 1990, 10 (06) : 2503 - 2512
  • [18] CONTACT OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE 1-INFECTED AND UNINFECTED CD4(+) T-LYMPHOCYTES IS HIGHLY CYTOLYTIC FOR BOTH CELLS
    HEINKELEIN, M
    SOPPER, S
    JASSOY, C
    [J]. JOURNAL OF VIROLOGY, 1995, 69 (11) : 6925 - 6931
  • [19] Activity of human immunodeficiency virus type 1 promoter TAR regions and tat1 genes derived from individuals with different rates of disease progression
    Kirchhoff, F
    Greenough, TC
    Hamacher, M
    Sullivan, JL
    Desrosiers, RC
    [J]. VIROLOGY, 1997, 232 (02) : 319 - 331
  • [20] RAF-1 PROTEIN-KINASE IS REQUIRED FOR GROWTH OF INDUCED NIH/3T3 CELLS
    KOLCH, W
    HEIDECKER, G
    LLOYD, P
    RAPP, UR
    [J]. NATURE, 1991, 349 (6308) : 426 - 428