Comparative transcriptomic analysis of Rickettsia conorii during in vitro infection of human and tick host cells.
Comparative transcriptomic analysis of Rickettsia conorii during in vitro infection of human and tick host cells.
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DOI：
10.1186/s12864-020-07077-w
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发表时间：
2020-09-25
期刊：
影响因子：
4.4
通讯作者：
Sahni SK
中科院分区：
文献类型：
--
作者：
Narra HP;Sahni A;Alsing J;Schroeder CLC;Golovko G;Nia AM;Fofanov Y;Khanipov K;Sahni SK
斑点热群致病立克次体是节肢动物传播的专性细胞内细菌，在哺乳动物宿主体内表现出对宿主微血管内皮细胞的优势趋向性，导致主要归因于内皮细胞损伤或功能障碍的疾病表现。虽然已知立克次体是通过基因组减少进行进化的，但这些病原体调节其转录组以确保在扁虱载体中存活并通过经卵巢/经生殖道传播维持生存的机制仍不清楚，而不是它们在人类宿主中引起衰弱感染的能力。在这项研究中，我们比较了立克次体sRNA组/转录组的表达谱，并确定了康氏立克次体转录本在体外感染人和蜱类宿主细胞过程中的转录起点(TSS)。我们对感染康氏杆菌的美洲Amblyomma americanum AAE2细胞和人微血管内皮细胞(HMECs)的总RNA进行了深度测序。对转录本进行链特异性RNA测序，发现32个小RNAs(Rcsr‘s)在Tick细胞感染过程中优先表达，定量RT-PCR证实RcsR61、SR71和sR74有表达。有趣的是，在AAE2细胞和HMEC中，共有305个和132个康氏杆菌编码基因被差异上调(>； 2倍)。此外，通过终止子5‘-磷酸依赖的核酸外切酶处理对初级转录本进行浓缩，获得了3903个和2555个转录起始位点(TSS)，其中包括在感染森林立克和人类宿主细胞期间分别鉴定出214个和181个初级TSS。75个编码基因在不同的寄主环境中表现出不同的SSs。最后，我们还观察到6S RNA在体外宿主-病原体和病媒-病原体相互作用中的差异表达，表明这种非编码RNA在依赖于支持宿主的生态位调控立克次体转录组中发挥重要作用。综上所述，本研究结果证实了新的Rcsr基因在水稻纹枯病菌中的存在，首次揭示了编码转录本的差异表达和依赖于寄主生态位的交替转录起始点的利用，并暗示了6S RNA在宿主-病原体-载体三方相互作用中对编码转录组的调控作用。
Pathogenic Rickettsia species belonging to the spotted fever group are arthropod-borne, obligate intracellular bacteria which exhibit preferential tropism for host microvascular endothelium in the mammalian hosts, resulting in disease manifestations attributed primarily to endothelial damage or dysfunction. Although rickettsiae are known to undergo evolution through genomic reduction, the mechanisms by which these pathogens regulate their transcriptome to ensure survival in tick vectors and maintenance by transovarial/transstadial transmission, in contrast to their ability to cause debilitating infections in human hosts remain unknown. In this study, we compare the expression profiles of rickettsial sRNAome/transcriptome and determine the transcriptional start sites (TSSs) of R. conorii transcripts during in vitro infection of human and tick host cells. We performed deep sequencing on total RNA from Amblyomma americanum AAE2 cells and human microvascular endothelial cells (HMECs) infected with R. conorii. Strand-specific RNA sequencing of R. conorii transcripts revealed the expression 32 small RNAs (Rc_sR’s), which were preferentially expressed above the limit of detection during tick cell infection, and confirmed the expression of Rc_sR61, sR71, and sR74 by quantitative RT-PCR. Intriguingly, a total of 305 and 132 R. conorii coding genes were differentially upregulated (> 2-fold) in AAE2 cells and HMECs, respectively. Further, enrichment for primary transcripts by treatment with Terminator 5′-Phosphate-dependent Exonuclease resulted in the identification of 3903 and 2555 transcription start sites (TSSs), including 214 and 181 primary TSSs in R. conorii during the infection to tick and human host cells, respectively. Seventy-five coding genes exhibited different TSSs depending on the host environment. Finally, we also observed differential expression of 6S RNA during host-pathogen and vector-pathogen interactions in vitro, implicating an important role for this noncoding RNA in the regulation of rickettsial transcriptome depending on the supportive host niche. In sum, the findings of this study authenticate the presence of novel Rc_sR’s in R. conorii, reveal the first evidence for differential expression of coding transcripts and utilization of alternate transcriptional start sites depending on the host niche, and implicate a role for 6S RNA in the regulation of coding transcriptome during tripartite host-pathogen-vector interactions.
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影响因子：
3.7
作者：
Galletti MF;Fujita A;Nishiyama MY Jr;Malossi CD;Pinter A;Soares JF;Daffre S;Labruna MB;Fogaça AC
通讯作者：
Fogaça AC
影响因子：
3.1
作者：
Allmond, LR;Karaca, TJ;Sawa, T
通讯作者：
Sawa, T
影响因子：
3.2
作者：
Beyer, Andrea R.;VieBrock, Lauren;Carlyon, Jason A.
通讯作者：
Carlyon, Jason A.
影响因子：
4.2
作者：
Dreher-Lesnick SM;Ceraul SM;Rahman MS;Azad AF
通讯作者：
Azad AF
影响因子：
3.7
作者：
Audoly G;Vincentelli R;Edouard S;Georgiades K;Mediannikov O;Gimenez G;Socolovschi C;Mège JL;Cambillau C;Raoult D
通讯作者：
Raoult D