The Anaplasma phagocytophilum PleC Histidine Kinase and PleD Diguanylate Cyclase Two-Component System and Role of Cyclic Di-GMP in Host Cell Infection

被引:46
|
作者
Lai, Tzung-Huei
Kumagai, Yumi
Hyodo, Mamoru [2 ]
Hayakawa, Yoshihiro [2 ]
Rikihisa, Yasuko [1 ]
机构
[1] Ohio State Univ, Dept Vet Biosci, Coll Vet Med, Columbus, OH 43210 USA
[2] Nagoya Univ, Bioorgan Chem Lab, Grad Sch Informat Sci, Nagoya, Aichi 4648601, Japan
基金
美国国家卫生研究院;
关键词
HUMAN GRANULOCYTIC EHRLICHIOSIS; CAULOBACTER-CRESCENTUS; RESPONSE REGULATOR; ENDOTHELIAL-CELLS; UNITED-STATES; PROTEINS; DOMAIN; AGENT; CHAFFEENSIS; TRANSITION;
D O I
10.1128/JB.01218-08
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Anaplasma phagocytophilum, the etiologic agent of human granulocytic anaplasmosis (HGA), has genes predicted to encode three sensor kinases, one of which is annotated PleC, and three response regulators, one of which is PleD. Prior to this study, the roles of PleC and PleD in the obligatory intracellular parasitism of A. phagocytophilum and their biochemical activities were unknown. The present study illustrates the relevance of these factors by demonstrating that both pleC and pleD were expressed in an HGA patient. During A. phagocytophilum development in human promyelocytic HL-60 cells, PleC and PleD were synchronously up-regulated at the exponential growth stage and downregulated prior to extracellular release. A recombinant PleC kinase domain (rPleCHKD) has histidine kinase activity; no activity was observed when the conserved site of phosphorylation was replaced with alanine. A recombinant PleD (rPleD) has autokinase activity using phosphorylated rPleCHKD as the phosphoryl donor but not with two other recombinant histidine kinases. rPleCHKD could not serve as the phosphoryl donor for a mutant rPleD ( with a conserved aspartic acid, the site of phosphorylation, replaced by alanine) or two other A. phagocytophilum recombinant response regulators. rPleD had diguanylate cyclase activity to generate cyclic ( c) di-GMP from GTP in vitro. UV cross-linking of A. phagocytophilum lysate with c-di-[32P] GMP detected an similar to 47-kDa endogenous protein, presumably c-di- GMP downstream receptor. A new hydrophobic c-di- GMP derivative, 2'-O-di(tert-butyldimethylsilyl)-c-di-GMP, inhibited A. phagocytophilum infection in HL-60 cells. Our results suggest that the two-component PleC-PleD system is a diguanylate cyclase and that a c-di-GMP-receptor complex regulates A. phagocytophilum intracellular infection.
引用
收藏
页码:693 / 700
页数:8
相关论文
共 4 条
  • [1] A Cyclic di-GMP-binding Adaptor Protein Interacts with Histidine Kinase to Regulate Two-component Signaling
    Xu, Linghui
    Venkataramani, Prabhadevi
    Ding, Yichen
    Liu, Yang
    Deng, Yinyue
    Yong, Grace Lisi
    Xin, Lingyi
    Ye, Ruijuan
    Zhang, Lianhui
    Yang, Liang
    Liang, Zhao-Xun
    JOURNAL OF BIOLOGICAL CHEMISTRY, 2016, 291 (31) : 16112 - 16123
  • [2] A Two-Component System That Modulates Cyclic di-GMP Metabolism Promotes Legionella pneumophila Differentiation and Viability in Low-Nutrient Conditions
    Hughes, Elisa D.
    Byrne, Brenda G.
    Swanson, Michele S.
    JOURNAL OF BACTERIOLOGY, 2019, 201 (17)
  • [3] The Two-Component System FleS/FleR Represses H1-T6SS via Cyclic di-GMP Signaling in Pseudomonas aeruginosa
    Zhou, Tian
    Huang, Jiahui
    Liu, Zhiqing
    Lin, Qiqi
    Xu, Zeling
    Zhang, Lian-Hui
    APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2022, 88 (02)
  • [4] The Atypical Two-component Sensor Kinase Lpl0330 from Legionella pneumophila Controls the Bifunctional Diguanylate Cyclase-Phosphodiesterase Lpl0329 to Modulate Bis-(3′-5′)-cyclic Dimeric GMP Synthesis
    Levet-Paulo, Melanie
    Lazzaroni, Jean-Claude
    Gilbert, Christophe
    Atlan, Daniele
    Doublet, Patricia
    Vianney, Anne
    JOURNAL OF BIOLOGICAL CHEMISTRY, 2011, 286 (36) : 31136 - 31144