Molecular characterization and functional analysis of Eimeria tenella ankyrin repeat-containing protein

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2024-05-10 DOI:10.1016/j.ejop.2024.126089

Huilin Guo , Qiping Zhao , Haixia Wang, Shunhai Zhu, Hui Dong, Xinrui Xie, Lihui Wang, Lang Chen, Hongyu Han

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Abstract

Chicken coccidiosis causes disastrous losses to the poultry industry all over the world. Eimeria tenella is the most prevalent of these disease-causing species. Our former RNA-seq indicated that E. tenella ankyrin repeat-containing protein (EtANK) was expressed differently between drug-sensitive (DS) and drug-resistant strains. In this study, we cloned EtANK and analyzed its translational and transcriptional levels using quantitative real-time PCR (qPCR) and western blotting. The data showed that EtANK was significantly upregulated in diclazuril-resistant (DZR) strain and maduramicin-resistant (MRR) strain compared with the drug-sensitive (DS) strain. In addition, the transcription levels in the DZR strains isolated from the field were higher than in the DS strain. The translation levels of EtANK were higher in unsporulated oocysts (UO) than in sporozoites (SZ), sporulated oocysts (SO), or second-generation merozoites (SM), and the protein levels in SM were significantly higher than in UO, SO, and SZ. The results of the indirect immunofluorescence localization showed that the protein was distributed mainly at the anterior region of SZ and on the surface and in the cytoplasm of SM. The fluorescence intensity increased further with its development in vitro. An anti-rEtANK polyclonal antibody inhibited the invasive ability of E. tenella in DF-1 cells. These results showed that EtANK may be related to host cell invasion, required for the parasite’s growth in the host, and may be involved in the development of E. tenella resistance to some drugs.

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天牛埃默氏菌含 ankyrin 重复蛋白的分子特征和功能分析

鸡球虫病给世界各地的家禽业造成了灾难性的损失。天牛埃默氏菌是这些致病菌中最常见的一种。我们以前的 RNA-seq 研究表明，天牛埃默氏菌含 ankyrin 重复蛋白（EtANK）在药物敏感株（DS）和耐药株之间的表达量不同。在本研究中，我们克隆了 EtANK，并利用实时定量 PCR（qPCR）和 Western 印迹分析了其转译和转录水平。数据显示，与药物敏感株（DS）相比，EtANK在地克珠利耐药株（DZR）和马杜霉素耐药株（MRR）中明显上调。此外，从田间分离的 DZR 菌株的转录水平高于 DS 菌株。未孢子化卵囊（UO）中的EtANK翻译水平高于孢子化卵囊（SZ）、孢子化卵囊（SO）或第二代子囊虫（SM），SM中的蛋白质水平也明显高于UO、SO和SZ。间接免疫荧光定位的结果表明，该蛋白主要分布在 SZ 的前部、SM 的表面和细胞质中。随着体外发育，荧光强度进一步增加。抗 EtANK 多克隆抗体抑制了天牛对 DF-1 细胞的侵袭能力。这些结果表明，EtANK可能与宿主细胞侵袭有关，是寄生虫在宿主体内生长所必需的，并可能参与了天牛血吸虫对某些药物产生抗药性的过程。

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

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.