Shen Liheng, Xiong Xiaobing, Xu Zixiang, Liu Yingli, Sheng Yan, Sheng Xin
Department of Biochemistry, Zunyi Medical University, Zunyi, China.
Laboratory of Basic Medical Morphology, Zunyi Medical University, Zunyi, China.
Front Microbiol. 2025 May 13;16:1486189. doi: 10.3389/fmicb.2025.1486189. eCollection 2025.
Ciliates serve as pivotal model organisms for investigating the protein composition and regulatory mechanisms underlying cellular processes. This study systematically explores the structural and functional characteristics of cilia-associated genes in , aiming to elucidate their roles in ciliogenesis and cilia-related pathways.
The macronuclear genome of was sequenced using next-generation sequencing (NGS). Cilia-associated genes were identified via BLASTP analysis against homologs in hypotrich ciliates (, , and ). Functional annotations, including Non-Redundant (NR) classification, Pfam domain prediction, and Gene Ontology (GO) enrichment, were performed. Pathway enrichment analysis identified associated metabolic and signaling pathways. Experimental validation included quantitative polymerase chain reaction (QPCR) of cilia-related gene families, RNA interference (RNAi) targeting ARL2BP and DYNLRB2, and immunofluorescence staining to assess microtubule arrangement.
A total of 418 cilia-associated genes were identified, with 44 conserved across the four hypotrich ciliate species. Functional categorization revealed kinases, dyneins, tubulins, and autophagy-related proteins. Pfam annotations predicted three conserved domains. GO terms were enriched in tubulin binding, cilia assembly, and microtubule-based movement. Pathway analysis implicated these genes in adenine ribonucleotide biosynthesis, glycolysis/gluconeogenesis, Wnt, and AMP-activated protein kinase (AMPK) signaling. QPCR showed significant downregulation of cilia-related proteins during mitosis. RNAi of ARL2BP and DYNLRB2 increased mortality, reduced motility, and disrupted cortical microtubule organization via immunofluorescence. Thirty-nine hub genes were strongly linked to ciliopathies.
Cilia-associated genes in are integral to DNA replication, energy metabolism, intercellular communication, and morphogenesis. The conserved hub genes associated with ciliopathies suggest evolutionary preservation of ciliogenesis regulation. ARL2BP and DYNLRB2 are functional important in ciliary dynamics and structural integrity. This study provides crucial insights into the roles of cilia-associated genes in ciliates, advancing understanding of ciliogenesis mechanisms and their implications for ciliopathy research.
纤毛虫是研究细胞过程中蛋白质组成和调控机制的关键模式生物。本研究系统地探索了[具体物种]中纤毛相关基因的结构和功能特征，旨在阐明它们在纤毛发生和纤毛相关途径中的作用。
使用下一代测序（NGS）对[具体物种]的大核基因组进行测序。通过针对低等毛口类纤毛虫（[具体物种1]、[具体物种2]和[具体物种3]）中的同源物进行BLASTP分析来鉴定纤毛相关基因。进行了功能注释，包括非冗余（NR）分类、Pfam结构域预测和基因本体（GO）富集分析。通路富集分析确定了相关的代谢和信号通路。实验验证包括对纤毛相关基因家族进行定量聚合酶链反应（QPCR）、针对ARL2BP和DYNLRB2的RNA干扰（RNAi）以及免疫荧光染色以评估微管排列。
共鉴定出418个纤毛相关基因，其中44个在四种低等毛口类纤毛虫物种中保守。功能分类显示有激酶、动力蛋白、微管蛋白和自噬相关蛋白。Pfam注释预测了三个保守结构域。GO术语富集在微管蛋白结合、纤毛组装和基于微管的运动方面。通路分析表明这些基因参与腺嘌呤核糖核苷酸生物合成、糖酵解/糖异生、Wnt和AMP激活的蛋白激酶（AMPK）信号传导。QPCR显示有丝分裂期间纤毛相关蛋白显著下调。ARL2BP和DYNLRB2的RNAi增加了死亡率，降低了运动能力，并通过免疫荧光破坏了皮层微管组织。39个枢纽基因与纤毛病密切相关。
[具体物种]中的纤毛相关基因对于DNA复制、能量代谢、细胞间通讯和形态发生至关重要。与纤毛病相关的保守枢纽基因表明纤毛发生调控在进化上具有保守性。ARL2BP和DYNLRB2在纤毛动力学和结构完整性方面具有重要功能。本研究为纤毛虫中纤毛相关基因的作用提供了关键见解，推进了对纤毛发生机制及其对纤毛病研究意义的理解。
