Exploring Tr1-regulated transcription networks underpinning adaptation of pathogenic Anaplasma to the tick host

探索 Tr1 调节的转录网络支持致病性无形体对蜱宿主的适应

• 批准号: 10727435
• 负责人: Jason Michael Park
• 金额: $ 21.54万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10727435
• 关键词: Anaplasma; Anaplasma phagocytophilum; Animals; Architecture; Arthropod Vectors; Arthropods; Attenuated; Bacteria; Binding; Bioinformatics; Biology; Body Temperature; Cell Culture Techniques; Cell Survival; Cells; Cues; DNA; DNA Binding; DNA Binding Domain; DNA Sequence; DNA-Binding Proteins; Dependence; Development; Disease; Disparate; Disparity; Environment; Evolution; Face; Family; Gene Expression; Genes; Genetic Transcription; Genome; Genomics; Growth; Helix-Turn-Helix Motifs; Homologous Gene; Human; Human Cell Line; Immunologics; Infection; Intervention; Lead; Life Cycle Stages; Life Style; Livestock; Mammalian Cell; Mammals; Measures; Modeling; Mus; Mutate; Mutation; Nutrient availability; Operon; Pathogenicity; Pattern; Physiological; Proteins; Regulon; Repression; Research; Response Elements; Rickettsia; Role; Side; Stimulus; Structural Models; Testing; Tick-Borne Diseases; Ticks; Transcriptional Regulation; United States; Vector-transmitted infectious disease; Work; Xenobiotics; disease transmission; gene network; genetic regulatory protein; innovation; mutant; pathogen; pathogenic bacteria; physical property; pressure; response; tick-borne; transcription factor; transcriptome; transmission process; vector; vector transmission; vector-borne; vector-borne pathogen

Project Summary Tick-borne diseases are on the increase and are responsible for nearly all of the vector-transmitted disease in the US. Vector-borne pathogens face the dual challenge of adaptation to two very different host environments: the arthropod vector and the mammalian host. Ticks contain distinct physiological cues including disparities in body temperature, nutrient availability, physiological architecture, and unique immunological pressures. Once in the tick, A. phagocytophilum must further adapt to construct a replicative niche within the arthropod’s cells. In response to the tick environment, A. phagocytophilum differentially transcribes 41% of its genes when infecting tick cells in comparison to mammalian cell culture. It is not known what controls this extensive reprogramming or how it facilitates A. phagocytophilum adaptation to the tick. One predicted transcription factor, tr1, displays the highest tick-specific expression of all A. phagocytophilum genes. Our structural modeling identifies Tr1 as a homo-dimeric helix-turn-helix DNA binding protein in the xenobiotic response element family of transcription factors. Disruption of tr1 by transposon insertion had no impact on bacterial burden in mice or growth in human cell lines. However, A. phagocytophilum lacking tr1 was greatly attenuated for acquisition by ticks and is unable to survival in tick cells. Given the importance of Tr1 for survival in the arthropod vector and its predicted role as a transcriptional switch, we hypothesize that: Tr1 operates as a master regulator for tick adaptation by orchestrating the expression of tick-specific gene networks. In this study we will identify the DNA sequences bound by Tr1 and its mode of transcription regulation. Further, we will identify how Tr1 contributes to completion of the tick cell infection cycle and measure how Tr1 remodels the A. phagocytophilum transcriptome during tick cell infection. Revealing how A. phagocytophilum adapts to infect tick cells will open the door to development of vector targeted interventions to reduce transmissibility of the pathogen.

项目摘要 蜱传疾病正在增加，几乎是所有病媒传播疾病的原因 在美国.媒介传播的病原体面临着适应两种截然不同的宿主的双重挑战 环境：节肢动物载体和哺乳动物宿主。蜱虫包含不同的生理线索 包括体温、营养物质供应、生理结构和独特的 免疫压力一旦在滴答声中，A。嗜吞噬细胞菌必须进一步适应， 在节肢动物细胞内的小生境。A.差异嗜吞噬细胞菌 与哺乳动物细胞培养物相比，在感染蜱细胞时转录41%的基因。目前还不知道 是什么控制了这种广泛的重新编程，或者它是如何促进A.嗜吞噬细胞菌对蜱的适应一 预测的转录因子tr 1在所有A.嗜吞噬细胞菌基因 我们的结构模型鉴定Tr 1为异生素中的同源二聚螺旋-转角-螺旋DNA结合蛋白。 转录因子应答元件家族。转座子插入对tr 1的破坏没有影响， 小鼠中的细菌负荷或人细胞系中的生长。然而，A.缺乏tr 1的嗜吞噬细胞 对于蜱的获取是减毒的，并且不能在蜱细胞中存活。鉴于Tr 1对生存的重要性 在节肢动物载体及其作为转录开关的预测作用中，我们假设：Tr 1作为一个转录开关， 通过协调蜱虫特异性基因网络的表达，成为蜱虫适应的主要调节因子。本研究 我们将鉴定Tr 1结合的DNA序列及其转录调节模式。此外，我们将 确定Tr 1如何促进蜱细胞感染周期的完成，并测量Tr 1如何重塑A. 在蜱细胞感染期间嗜吞噬细胞菌转录组。揭示了A.嗜吞噬细胞菌适应感染 蜱细胞将打开大门，发展载体靶向干预措施，以减少传播的 病原体