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Methionine sulfoxide reductase A (MsrA) modulates cells and protects against Mycoplasma genitalium induced cytotoxicity.
Free Radical Biology and Medicine ( IF 7.1 ) Pub Date : 2020-03-25 , DOI: 10.1016/j.freeradbiomed.2020.03.019
Kishore Das 1 , Omar Garnica 1 , Javier Flores 2 , Subramanian Dhandayuthapani 2
Affiliation  

Methionine sulfoxide reductase A (MsrA) is a ubiquitous antioxidant repair enzyme which specifically reduces the oxidized methionine (Met-O) in proteins to methionine (Met). Previous studies have shown that lack of or overexpression of MsrA in cells affects the function of proteins and can lead to altered cellular processes. Interestingly, some pathogenic bacteria secrete and/or carry MsrA on their surface, suggesting some key roles for this enzyme in the modulation of host cellular processes. Therefore, we investigated how exogenously added MsrA affects the ability of the host cells in combating infection by using an in vitroMycoplasma genitalium cytotoxicity model. HeLa cells pretreated with MsrA and infected with M. genitalium showed significantly lower necrosis (cytotoxicity) than untreated cells infected with M. genitalium. Intriguingly, necrotic cell death pathway specific real time RT-PCR revealed that M. genitalium infection upregulates the expression of the TNF gene in HeLa cells and that MsrA pretreatment of the cells downregulates its expression significantly. Consistent with this, enzyme linked immunosorbent assay (ELISA) results showed that HeLa cells pretreated with MsrA secreted reduced levels of TNF-α following M. genitalium infection. Also, our study demonstrates that MsrA treatment of cells affects the phosphorylation status of transcriptional regulators such as NF-кB, JNK and p53 that regulate different cytokines. Further, fluorescent microscopy showed the cellular uptake of exogenously added MsrA fused with red fluorescent protein (MsrA-RFP). Altogether, our results suggest that secreted MsrA may help pathogens to modulate host cellular processes.

中文翻译:

蛋氨酸亚砜还原酶 A (MsrA) 调节细胞并防止生殖支原体诱导的细胞毒性。

蛋氨酸亚砜还原酶 A (MsrA) 是一种普遍存在的抗氧化修复酶,可特异性地将蛋白质中的氧化蛋氨酸 (Met-O) 还原为蛋氨酸 (Met)。先前的研究表明,细胞中 MsrA 的缺乏或过度表达会影响蛋白质的功能,并可能导致细胞过程的改变。有趣的是,一些病原菌在其表面分泌和/或携带 MsrA,这表明这种酶在调节宿主细胞过程中具有一些关键作用。因此,我们使用体外生殖支原体细胞毒性模型研究了外源性添加的 MsrA 如何影响宿主细胞对抗感染的能力。用 MsrA 预处理并感染 M. genitalium 的 HeLa 细胞的坏死(细胞毒性)明显低于感染 M. genitalium 的未处理细胞。耐人寻味的是,坏死细胞死亡通路特异性实时 RT-PCR 显示,生殖器分枝杆菌感染上调了 HeLa 细胞中 TNF 基因的表达,而细胞的 MsrA 预处理显着下调了其表达。与此一致,酶联免疫吸附试验 (ELISA) 结果表明,用 MsrA 预处理的 HeLa 细胞在生殖支原体感染后分泌的 TNF-α 水平降低。此外,我们的研究表明,细胞的 MsrA 处理会影响转录调节因子的磷酸化状态,例如调节不同细胞因子的 NF-кB、JNK 和 p53。此外,荧光显微镜显示细胞吸收了与红色荧光蛋白 (MsrA-RFP) 融合的外源添加的 MsrA。总之,我们的结果表明分泌的 MsrA 可能有助于病原体调节宿主细胞过程。
更新日期:2020-03-26
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