An Ehrlichia chaffeensis tandem repeat protein interacts with multiple host targets involved in cell signaling, transcriptional regulation, and vesicle trafficking.
An Ehrlichia chaffeensis tandem repeat protein interacts with multiple host targets involved in cell signaling, transcriptional regulation, and vesicle trafficking.
复制标题
恰菲埃里希体串联重复蛋白与参与细胞信号传导、转录调控和囊泡运输的多个宿主靶标相互作用。
DOI：
10.1128/iai.00027-09
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发表时间：
2009
影响因子：
3.1
通讯作者：
McBride,JereW
中科院分区：
文献类型：
--
作者：
Wakeel,Abdul;Kuriakose,JeebaA;McBride,JereW
沙芬埃立体是一种专性的细胞内细菌，表现出单核吞噬细胞形成细胞质膜结合微菌落的倾向。为了在吞噬细胞内存活和复制，E。chaffenensis通过调节宿主细胞的许多过程来利用宿主细胞，但所涉及的卵泡效应蛋白尚不清楚。在这项研究中，我们确定p47是一种分泌的、差异表达的串联重复序列（TR）蛋白，与多种与细胞信号传导、转录调控和囊泡运输相关的宿主蛋白相互作用。酵母双杂交分析发现，p47与多梳群无名指5蛋白（PCGF5）、Src蛋白酪氨酸激酶FYN （FYN）、蛋白酪氨酸磷酸酶非受体2型（PTPN2）和腺苷酸环化酶相关蛋白1 （CAP1）相互作用。在转染了p47-绿色荧光融合蛋白（AcGFP1-p47）的HeLa细胞中，通过共免疫沉淀实验和共定位进一步证实了p47与这些蛋白的相互作用。此外，共聚焦显微镜显示表达p47的致密核（DC）埃氏体与PCGF5、FYN、PTPN2和CAP1共定位。含有TRs的氨基末端截断形式的p47仅与PCGF5相互作用，而不与FYN， PTPN2和CAP1相互作用，这表明参与这些相互作用的p47结构域存在差异。这些结果表明，p47参与了一个复杂的相互作用网络，涉及许多宿主细胞蛋白。此外，本研究还提供了新的见解，以了解DC ehrlichiae的分子和功能差异，以及参与促进ehrlichiae在单核吞噬细胞中存活的效应蛋白。
Ehrlichia chaffeensisis an obligately intracellular bacterium that exhibits tropism for mononuclear phagocytes forming cytoplasmic membrane-bound microcolonies called morulae. To survive and replicate within phagocytes,E. chaffeensisexploits the host cell by modulating a number of host cell processes, but the ehrlichial effector proteins involved are unknown. In this study, we determined that p47, a secreted, differentially expressed, tandem repeat (TR) protein, interacts with multiple host proteins associated with cell signaling, transcriptional regulation, and vesicle trafficking. Yeast two-hybrid analysis revealed that p47 interacts with polycomb group ring finger 5 (PCGF5) protein, Src protein tyrosine kinase FYN (FYN), protein tyrosine phosphatase non-receptor type 2 (PTPN2), and adenylate cyclase-associated protein 1 (CAP1). p47 interaction with these proteins was further confirmed by coimmunoprecipitation assays and colocalization in HeLa cells transfected with p47-green fluorescent fusion protein (AcGFP1-p47). Moreover, confocal microscopy demonstrated p47-expressing dense-cored (DC) ehrlichiae colocalized with PCGF5, FYN, PTPN2, and CAP1. An amino-terminally truncated form of p47 containing TRs interacted only with PCGF5 and not with FYN, PTPN2, and CAP1, indicating differences in p47 domains that are involved in these interactions. These results demonstrate that p47 is involved in a complex network of interactions involving numerous host cell proteins. Furthermore, this study provides a new insight into the molecular and functional distinction of DC ehrlichiae, as well as the effector proteins involved in facilitating ehrlichial survival in mononuclear phagocytes.