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首页> 外文期刊>Infection and immunity >Identification of Critical Host Mitochondrion-Associated Genes during Ehrlichia chaffeensis Infections
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Identification of Critical Host Mitochondrion-Associated Genes during Ehrlichia chaffeensis Infections

机译：恰菲埃里希氏菌感染过程中关键宿主线粒体相关基因的鉴定。

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Ehrlichia chaffeensis is an obligate intracellular bacterium that causes human monocytic ehrlichiosis (HME). To determine what host components are important for bacterial replication, we performed microarray analysis on Drosophila melanogaster S2 cells by comparing host gene transcript levels between permissive and nonpermissive conditions for E. chaffeensis growth. Five-hundred twenty-seven genes had increased transcript levels unique to permissive growth conditions 24 h postinfection. We screened adult flies that were mutants for several of the “permissive” genes for the ability to support Ehrlichia replication. Three additional D. melanogaster fly lines with putative mutations in pyrimidine metabolism were also tested. Ten fly lines carrying mutations in the genes CG6479, separation anxiety, chitinase 11, CG6364 (Uck2), CG6543 (Echs1), withered (whd), CG15881 (Ccdc58), CG14806 (Apop1), CG11875 (Nup37), and dumpy (dp) had increased resistance to infection with Ehrlichia. Analysis of RNA by quantitative real-time reverse transcription-PCR (qRT-PCR) confirmed that the bacterial load was decreased in these mutant flies compared to wild-type infected control flies. Seven of these genes (san, Cht11, Uck2, Echs1, whd, Ccdc58, and Apop1) encoded proteins that had mitochondrial functions or could be associated with proteins with mitochondrial functions. Treatment of THP-1 cells with double-stranded RNA to silence the human UCK2 gene indicates that the disruption of the uridine-cytidine kinase affects E. chaffeensis replication in human macrophages. Experiments with cyclopentenyl cytosine (CPEC), a CTP synthetase inhibitor and cytosine, suggest that the nucleotide salvage pathway is essential for E. chaffeensis replication and that it may be important for the provision of CTP, uridine, and cytidine nucleotides.
机译：恰菲埃里希氏菌是专一的细胞内细菌，可引起人单核细胞埃希氏菌病（HME）。为了确定哪些宿主成分对于细菌复制很重要，我们通过比较果蝇大肠杆菌生长的允许和不允许条件之间的宿主基因转录水平，对果蝇S2细胞进行了微阵列分析。 257个基因在感染后24小时内具有允许的生长条件所独有的转录水平。我们筛选了成年蝇，这些蝇是支持埃希氏菌复制能力的几个“允许”基因的突变体。还测试了在嘧啶代谢中具有推定突变的另外三个黑腹果蝇蝇系。十条蝇线携带 CG6479 ，分离焦虑，几丁质酶11 ， CG6364 （ Uck2 ）， CG6543 （ Echs1 ），枯萎的（ whd ）， CG15881 （ Ccdc58 ）， CG14806 （ Apop1 ）， CG11875 （ Nup37 ）和 dumpy < / em>（ dp ）对埃希氏菌感染的抵抗力增强。通过定量实时逆转录PCR（qRT-PCR）对RNA的分析证实，与野生型感染的对照果蝇相比，这些突变果蝇的细菌载量降低了。这些基因中的七个（ san ， Cht11 ， Uck2 ， Echs1 ， whd ， C cdc58 和 Apop1 ）编码的蛋白具有线粒体功能，或者可能与具有线粒体功能的蛋白相关。用双链RNA处理THP-1细胞以使人 UCK2 基因沉默，这表明尿苷-胞苷激酶的破坏会影响人巨噬细胞中恰菲酵母的复制。用CTP合成酶抑制剂环戊烯基胞嘧啶（CPEC）和胞嘧啶进行的实验表明，核苷酸挽救途径对于Chaffeensis大肠杆菌的复制是必不可少的，并且对于CTP，尿苷和胞苷核苷酸的提供可能很重要。

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