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从秀丽隐杆线虫中负载的 miRNA 的细胞类型特异性分析揭示了 Argonaute 加载的空间和时间灵活性。

Cell-type-specific profiling of loaded miRNAs from Caenorhabditis elegans reveals spatial and temporal flexibility in Argonaute loading.

机构信息

Clem Jones Centre for Ageing Dementia Research, Queensland Brain Institute, The University of Queensland, Brisbane, Australia.

Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, Brisbane, Australia.

出版信息

Nat Commun. 2021 Apr 13;12(1):2194. doi: 10.1038/s41467-021-22503-7.

DOI:10.1038/s41467-021-22503-7
PMID:33850152
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8044110/
Abstract

Multicellularity has coincided with the evolution of microRNAs (miRNAs), small regulatory RNAs that are integrated into cellular differentiation and homeostatic gene-regulatory networks. However, the regulatory mechanisms underpinning miRNA activity have remained largely obscured because of the precise, and thus difficult to access, cellular contexts under which they operate. To resolve these, we have generated a genome-wide map of active miRNAs in Caenorhabditis elegans by revealing cell-type-specific patterns of miRNAs loaded into Argonaute (AGO) silencing complexes. Epitope-labelled AGO proteins were selectively expressed and immunoprecipitated from three distinct tissue types and associated miRNAs sequenced. In addition to providing information on biological function, we define adaptable miRNA:AGO interactions with single-cell-type and AGO-specific resolution. We demonstrate spatial and temporal dynamicism, flexibility of miRNA loading, and suggest miRNA regulatory mechanisms via AGO selectivity in different tissues and during ageing. Additionally, we resolve widespread changes in AGO-regulated gene expression by analysing translatomes specifically in neurons.

摘要

多细胞生物的出现与 microRNAs（miRNAs）的进化同时发生，miRNAs 是一种小型调控 RNA，整合到细胞分化和体内平衡的基因调控网络中。然而，由于它们作用的细胞环境精确而难以接近，miRNA 活性的调控机制在很大程度上仍未被揭示。为了解决这个问题，我们通过揭示 Argonaute（AGO）沉默复合物中加载的 miRNA 的细胞类型特异性模式，生成了秀丽隐杆线虫中全基因组活跃 miRNA 的图谱。通过选择性表达和免疫沉淀三种不同组织类型的带有表位标记的 AGO 蛋白，并对相关的 miRNA 进行测序。除了提供有关生物学功能的信息外，我们还以单细胞类型和 AGO 特异性分辨率定义了适应性 miRNA：AGO 相互作用。我们证明了 miRNA 加载的空间和时间动态性、灵活性，并通过不同组织和衰老过程中 AGO 的选择性提出了 miRNA 调控机制的假设。此外，我们通过专门分析神经元中的翻译组，解决了AGO 调控基因表达的广泛变化问题。

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