A hydrophobic binding surface on the human immunodeficiency virus type 1 Nef core is critical for association with p21-activated kinase 2

被引:45
作者
Agopian, K
Wei, BL
Garcia, JV
Gabuzda, D
机构
[1] Harvard Univ, Sch Med, Dana Farber Canc Inst, Dept Canc Immunol & AIDS, Boston, MA 02115 USA
[2] Harvard Univ, Sch Med, Dept Neurol, Boston, MA 02115 USA
[3] Univ Texas, SW Med Ctr, Dept Internal Med, Div Infect Dis, Dallas, TX 75390 USA
关键词
D O I
10.1128/JVI.80.6.3050-3061.2006
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The interaction of human immunodeficiency virus type 1 (HIV-1) Nef with p21-activated kinase 2 (Pak2) has been proposed to play an important role in T-cell activation and disease progression during viral infection. However, the mechanism by which Nef activates Pak2 is poorly understood. Mutations in most Nef motifs previously reported to be required for Pak2 activation (G(2), PxxP(72), and RR105) also affect other Nef functions, such as CD4 or major histocompatibility complex class I (MHC-I) downregulation. To better understand Nef interactions with Pak2, we performed mutational analysis of three primary HIV-1 Nef clones that exhibited similar capacities for downregulation of CD4 and MHC-I but variable abilities to associate with activated Pak2. Our results demonstrate that Nef amino acids at positions 85, 89, 187, 188, and 191 (L, H, S, R, and F in the clade B consensus, respectively) are critical for Pak2 association. Mutation of these Nef residues dramatically altered association with Pak2 without affecting Nef expression levels or CD4 and MHC-I downregulation. Furthermore, compensation occurred at positions 89 and 191 when both amino acids were substituted. Since residues 85, 89, 187, 188, and 191 cluster on the surface of the Nef core domain in a region distinct from the dimerization and SH3-binding domains, we propose that these Nef residues form part of a unique binding surface specifically involved in association with Pak2. This binding surface includes exposed and recessed hydrophobic residues and may participate in an as-yet-unidentified protein-protein interaction to facilitate Pak2 activation.
引用
收藏
页码:3050 / 3061
页数:12
相关论文
共 67 条
  • [1] A role for natural simian immunodeficiency virus and human immunodeficiency virus type 1 Nef alleles in lymphocyte activation
    Alexander, L
    Du, ZJ
    Rosenzweig, M
    Jung, JU
    Desrosiers, RC
    [J]. JOURNAL OF VIROLOGY, 1997, 71 (08) : 6094 - 6099
  • [2] The crystal structure of HIV-1 Nef protein bound to the Fyn kinase SH3 domain suggests a role for this complex in altered T cell receptor signaling
    Arold, S
    Franken, P
    Strub, MP
    Hoh, F
    Benichou, S
    Benarous, R
    Dumas, C
    [J]. STRUCTURE, 1997, 5 (10) : 1361 - 1372
  • [3] Dynamic Nef and Nef dynamics: how structure could explain the complex activities of this small HIV protein
    Arold, ST
    Bauer, AS
    [J]. TRENDS IN BIOCHEMICAL SCIENCES, 2001, 26 (06) : 356 - 363
  • [4] Lentivirus Nef specifically activates Pak2
    Arora, VK
    Molina, RP
    Foster, JL
    Blakemore, JL
    Chernoff, J
    Fredericksen, BL
    Garcia, JV
    [J]. JOURNAL OF VIROLOGY, 2000, 74 (23) : 11081 - 11087
  • [5] PAK to the future
    Bagrodia, S
    Cerione, RA
    [J]. TRENDS IN CELL BIOLOGY, 1999, 9 (09) : 350 - 355
  • [6] HIV-1 NEF LEADS TO INHIBITION OR ACTIVATION OF T-CELLS DEPENDING ON ITS INTRACELLULAR-LOCALIZATION
    BAUR, AS
    SAWAI, ET
    DAZIN, P
    FANTL, WJ
    CHENGMAYER, C
    PETERLIN, BM
    [J]. IMMUNITY, 1994, 1 (05) : 373 - 384
  • [7] Bour S, 2000, Adv Pharmacol, V48, P75
  • [8] A dileucine motif in HIV-1 Nef acts as an internalization signal for CD4 downregulation and binds the AP-1 clathrin adaptor
    Bresnahan, PA
    Yonemoto, W
    Ferrell, S
    Williams-Herman, D
    Geleziunas, R
    Greene, WC
    [J]. CURRENT BIOLOGY, 1998, 8 (22) : 1235 - 1238
  • [9] Activation of the PAK-related kinase by human immunodeficiency virus type 1 Nef in primary human peripheral blood lymphocytes and macrophages leads to phosphorylation of a PIX-p95 complex
    Brown, A
    Wang, X
    Sawai, E
    Cheng-Mayer, C
    [J]. JOURNAL OF VIROLOGY, 1999, 73 (12) : 9899 - 9907
  • [10] Simian immunodeficiency virus containing mutations in N-terminal tyrosine residues and in the PxxP motif in Nef replicates efficiently in rhesus macaques
    Carl, S
    Iafrate, AJ
    Lang, SM
    Stolte, N
    Stahl-Hennig, C
    Mätz-Rensing, K
    Fuchs, D
    Skowronski, J
    Kirchhoff, F
    [J]. JOURNAL OF VIROLOGY, 2000, 74 (09) : 4155 - 4164