旋毛虫蛋白酶体β5 亚基在生长、发育和寿命中的作用。

A proteasomal β5 subunit of Haemonchus contortus with a role in the growth, development and life span.

机构信息

State Key Laboratory of Agricultural Microbiology, Key Laboratory for the Development of Veterinary Products, Ministry of Agriculture College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, Hubei Province, People's Republic of China.

Hubei Key Laboratory of Embryonic Stem Cell Research, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, 442000, Hubei Province, People's Republic of China.

出版信息

Parasit Vectors. 2023 Mar 15;16(1):100. doi: 10.1186/s13071-023-05676-6.

DOI:10.1186/s13071-023-05676-6

PMID:36922877

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10015785/

Abstract

BACKGROUND

The proteasome in eukaryotic cells can degrade a variety of proteins and plays an important role in regulating the cell cycle, cell survival and apoptosis. The proteasome receives much attention as a potential chemotherapeutic target for treatment of a variety of infectious parasitic diseases, but few studies of proteasomes have been done on parasitic nematodes.

METHODS

A proteasomal β5 subunit encoding gene (named Hc-pbs-5) and its inferred product (Hc-PBS-5) in Haemonchus contortus were identified and characterized in this study. Then, the transcriptional profiles and anatomical expression were studied using an integrated molecular approach. Finally, a specific proteasome inhibitor bortezomib (BTZ), together with RNA interference (RNAi), was employed to assess the function of Hc-PBS-5.

RESULTS

Bioinformatic analysis revealed that the coding sequence of Hc-pbs-5 was 855 bp long and encoded 284 amino acids (aa). The predicted protein (Hc-PBS-5) had core conservative sequences (65-250 aa) belonging to N-terminal nucleophile (Ntn) family of hydrolases. Real-time PCR results revealed that Hc-pbs-5 was continuously transcribed in eight developmental stages with higher levels at the infective third-stage larvae (L3s) and adult males of H. contortus. Immunohistochemical results revealed that Hc-PBS-5 was expressed in intestine, outer cuticle, muscle cells under the outer cuticle, cervical glands and seminal vesicles of male adults and also in intestine, outer cuticle, cervical glands, uterine wall, eggs and ovaries of female adults of H. contortus. BTZ could reduce proportions of egg hatching, and the fourth-stage larvae (L4s) developed from the exsheathed L3s (xL3s) of H. contortus. In addition, silencing Hc-pbs-5 by soaking the specific double-stranded RNA (dsRNA) could decrease the transcription of Hc-pbs-5 and result in fewer xL3s developing to L4s in vitro.

CONCLUSIONS

These results indicate that proteasomal β5 subunit plays an important role in the growth, development and life span of H. contortus.

摘要

背景

真核细胞中的蛋白酶体可以降解多种蛋白质，在调节细胞周期、细胞存活和细胞凋亡方面发挥着重要作用。蛋白酶体作为治疗多种传染性寄生虫病的潜在化疗靶点受到了广泛关注，但对寄生线虫的蛋白酶体研究甚少。

方法

本研究鉴定并描述了旋毛虫蛋白酶体β5 亚基编码基因（命名为 Hc-pbs-5）及其编码产物（Hc-PBS-5）。然后，采用综合分子方法研究了其转录谱和解剖表达。最后，采用特异性蛋白酶体抑制剂硼替佐米（BTZ）联合 RNA 干扰（RNAi）评估 Hc-PBS-5 的功能。

结果

生物信息学分析显示，Hc-pbs-5 的编码序列长 855bp，编码 284 个氨基酸（aa）。预测蛋白（Hc-PBS-5）具有核心保守序列（65-250aa），属于 N 端亲核（Ntn）家族水解酶。实时 PCR 结果显示，Hc-pbs-5 在 8 个发育阶段持续转录，在感染性 3 期幼虫（L3s）和旋毛虫雄性成虫中表达水平较高。免疫组化结果显示，Hc-PBS-5 在旋毛虫雄性成虫的肠、外皮层、外皮层下肌肉细胞、颈腺和精囊以及雌性成虫的肠、外皮层、颈腺、子宫壁、卵和卵巢中表达。BTZ 可降低孵化比例，并减少从脱鞘 L3s（xL3s）发育而来的第 4 期幼虫（L4s）。此外，通过浸泡特异性双链 RNA（dsRNA）沉默 Hc-pbs-5 可降低 Hc-pbs-5 的转录水平，导致体外 xL3s 发育成 L4s 的数量减少。