Biochemical characterization of ClpB and DnaK from Anaplasma phagocytophilum

IF 3.3 3区 生物学 Q3 CELL BIOLOGY
Chathurange B. Ranaweera , Sunitha Shiva , Swetha Madesh , Deepika Chauhan , Roman R. Ganta , Michal Zolkiewski
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Abstract

Anaplasma phagocytophilum is an intracellular tick-transmitted bacterial pathogen that infects neutrophils in mammals and causes granulocytic anaplasmosis. In this study, we investigated the molecular chaperones ClpB and DnaK from A. phagocytophilum. In Escherichia coli, ClpB cooperates with DnaK and its co-chaperones DnaJ and GrpE in ATP-dependent reactivation of aggregated proteins. Since ClpB is not produced in metazoans, it is a promising target for developing antimicrobial therapies, which generates interest in studies on that chaperone’s role in pathogenic bacteria. We found that ClpB and DnaK are transcriptionally upregulated in A. phagocytophilum 3–5 days after infection of human HL-60 and tick ISE6 cells, which suggests an essential role of the chaperones in supporting the pathogen’s intracellular life cycle. Multiple sequence alignments show that A. phagocytophilum ClpB and DnaK contain all structural domains that were identified in their previously studied orthologs from other bacteria. Both A. phagocytophilum ClpB and DnaK display ATPase activity, which is consistent with their participation in the ATP-dependent protein disaggregation system. However, despite a significant sequence similarity between the chaperones from A. phagocytophilum and those from E. coli, the former were not as effective as their E. coli orthologs during reactivation of aggregated proteins in vitro and in supporting the survival of E. coli cells under heat stress. We conclude that the A. phagocytophilum chaperones might have evolved with distinct biochemical properties to maintain the integrity of pathogenic proteins under unique stress conditions of an intracellular environment of host cells.

噬噬原虫 ClpB 和 DnaK 的生化特征。
噬细胞无形体(Anaplasma phagocytophilum)是一种细胞内蜱传细菌病原体,会感染哺乳动物的中性粒细胞并导致粒细胞无形体病。在这项研究中,我们研究了噬吞噬细胞嗜血杆菌的分子伴侣 ClpB 和 DnaK。在大肠杆菌中,ClpB 与 DnaK 及其辅助伴侣 DnaJ 和 GrpE 合作,在 ATP 依赖性条件下重新激活聚集蛋白。由于ClpB不在类人猿体内产生,因此它是开发抗菌疗法的一个有希望的目标,这引起了人们对该伴侣在病原菌中作用的研究兴趣。我们发现,噬细胞噬甲虫在感染人类 HL-60 细胞和蜱 ISE6 细胞 3-5 天后,ClpB 和 DnaK 转录上调,这表明伴侣蛋白在支持病原体的细胞内生命周期中发挥着重要作用。多重序列比对结果表明,噬细胞甲虫 ClpB 和 DnaK 含有之前研究的其他细菌同源物中发现的所有结构域。噬菌体 ClpB 和 DnaK 都具有 ATP 酶活性,这与它们参与 ATP 依赖性蛋白质分解系统是一致的。然而,尽管噬噬甲虫的伴侣蛋白与大肠杆菌的伴侣蛋白在序列上有很大的相似性,但前者在体外重新激活聚集蛋白和支持大肠杆菌细胞在热应激下存活的效果却不如大肠杆菌的同源物。我们的结论是,噬噬甲虫伴侣蛋白可能具有独特的生化特性,能在宿主细胞内环境的独特应激条件下维持致病蛋白的完整性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Cell Stress & Chaperones
Cell Stress & Chaperones 生物-细胞生物学
CiteScore
7.60
自引率
2.60%
发文量
59
审稿时长
6-12 weeks
期刊介绍: Cell Stress and Chaperones is an integrative journal that bridges the gap between laboratory model systems and natural populations. The journal captures the eclectic spirit of the cellular stress response field in a single, concentrated source of current information. Major emphasis is placed on the effects of climate change on individual species in the natural environment and their capacity to adapt. This emphasis expands our focus on stress biology and medicine by linking climate change effects to research on cellular stress responses of animals, micro-organisms and plants.
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