Longitudinal map of transcriptome changes in the Lyme pathogen Borrelia burgdorferi during tick-borne transmission.
Longitudinal map of transcriptome changes in the Lyme pathogen Borrelia burgdorferi during tick-borne transmission.
复制标题
作者：
Sapiro AL;Hayes BM;Volk RF;Zhang JY;Brooks DM;Martyn C;Radkov A;Zhao Z;Kinnersley M;Secor PR;Zaro BW;Chou S
伯氏疏螺旋体（Bb）是莱姆病的病原体，当它在蜱虫媒介和脊椎动物宿主之间循环时，适应了截然不同的环境。在蜱虫吸血过程中，Bb改变其基因表达，为脊椎动物感染做准备；然而，从技术上讲，捕捉蜱虫体内几天内发生的全部转录变化是具有挑战性的。我们开发了一种实验方法来富集Bb细胞，以纵向定义其在传血过程中在肩胛骨蜱的淋巴内的全局转录组景观。我们鉴定出192个Bb基因，在宿主附着后1至4天的血食过程中，它们的表达发生了实质性的变化。大多数上调的基因编码在细胞包膜上发现的蛋白质或功能未知的蛋白质，包括45种外表面脂蛋白嵌入不寻常的富含蛋白质的Bb外壳。由于这些蛋白可能促进Bb与宿主的相互作用，我们利用质谱法鉴定与Bb物理相关的候选蜱虫蛋白。本文提出的Bb富集方法以及体外Bb转录组和候选蜱虫相互作用蛋白为研究蜱虫在其独特传播周期阶段启动和传播的关键决定因素提供了资源。
Borrelia burgdorferi (Bb), the causative agent of Lyme disease, adapts to vastly different environments as it cycles between tick vector and vertebrate host. During a tick bloodmeal, Bb alters its gene expression to prepare for vertebrate infection; however, the full range of transcriptional changes that occur over several days inside of the tick are technically challenging to capture. We developed an experimental approach to enrich Bb cells to longitudinally define their global transcriptomic landscape inside nymphal Ixodes scapularis ticks during a transmitting bloodmeal. We identified 192 Bb genes that substantially change expression over the course of the bloodmeal from 1 to 4 days after host attachment. The majority of upregulated genes encode proteins found at the cell envelope or proteins of unknown function, including 45 outer surface lipoproteins embedded in the unusual protein-rich coat of Bb. As these proteins may facilitate Bb interactions with the host, we utilized mass spectrometry to identify candidate tick proteins that physically associate with Bb. The Bb enrichment methodology along with the ex vivo Bb transcriptomes and candidate tick interacting proteins presented here provide a resource to facilitate investigations into key determinants of Bb priming and transmission during the tick stage of its unique transmission cycle.