An RNAi screen for conserved kinases that enhance microRNA activity after dauer in Caenorhabditis elegans
An RNAi screen for conserved kinases that enhance microRNA activity after dauer in Caenorhabditis elegans
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DOI：
10.1093/g3journal/jkae007
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发表时间：
2024-02-01
影响因子：
2.6
通讯作者：
Karp，Xantha
中科院分区：
文献类型：
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作者：
Roka Pun，Himal;Karp，Xantha
在不断变化的环境中，基因调控对于维持体内平衡至关重要。有些动物会经历一个抗应激的滞育阶段，以承受在发育过程中遇到的恶劣环境条件。microRNA是滞育期间和滞育后调节基因表达的机制之一。microRNA通过microRNA诱导的沉默复合物的活性在转录后下调靶基因。Argonaute是核心microRNA诱导的沉默复合物蛋白，其结合microRNA和其他microRNA诱导的沉默复合物蛋白。秀丽隐杆线虫中的2种主要microRNA Argonaute是ALG-1和ALG-2，它们的功能部分冗余。alg-1[alg-1（0）]的缺失导致渗透性发育表型，包括外阴缺陷和下皮细胞中幼虫细胞程序的重复。然而，如果alg-1（0）动物经历一个称为dauer的滞育阶段，这些表型基本上就不存在了。在dauer过程中或之后，相关microRNA的水平并不高，这表明microRNA诱导的沉默复合物的活性可能在这种情况下增强。为了鉴定alg-1（0）突变体在dauer后无外阴缺陷发育所需的基因，我们对编码保守激酶的基因进行了RNAi筛选。我们专注于激酶，因为它们在调节microRNA诱导的沉默复合物活性中的已知作用。我们发现4个激酶编码基因air-2、bub-1、chk-1和nekl-3的RNAi敲低导致alg-1（0）突变体在dauer后出现外阴缺陷和退化表型，并且这些缺陷在analg-1（0）背景中比在野生型中更明显。我们的研究结果暗示这些激酶作为微RNA诱导的沉默复合物活性的潜在调节剂在发育过程中在C。优美的
Gene regulation in changing environments is critical for maintaining homeostasis. Some animals undergo a stress-resistant diapause stage to withstand harsh environmental conditions encountered during development. MicroRNAs are one mechanism for regulating gene expression during and after diapause. MicroRNAs downregulate target genes posttranscriptionally through the activity of the microRNA-induced silencing complex. Argonaute is the core microRNA-induced silencing complex protein that binds to both the microRNA and to other microRNA-induced silencing complex proteins. The 2 major microRNA Argonautes in theCaenorhabditis eleganssoma are ALG-1 and ALG-2, which function partially redundantly. Loss ofalg-1[alg-1(0)] causes penetrant developmental phenotypes including vulval defects and the reiteration of larval cell programs in hypodermal cells. However, these phenotypes are essentially absent ifalg-1(0)animals undergo a diapause stage called dauer. Levels of the relevant microRNAs are not higher during or after dauer, suggesting that activity of the microRNA-induced silencing complex may be enhanced in this context. To identify genes that are required foralg-1(0)mutants to develop without vulval defects after dauer, we performed an RNAi screen of genes encoding conserved kinases. We focused on kinases because of their known role in modulating microRNA-induced silencing complex activity. We found RNAi knockdown of 4 kinase-encoding genes,air-2,bub-1,chk-1, andnekl-3, caused vulval defects and reiterative phenotypes inalg-1(0)mutants after dauer, and that these defects were more penetrant in analg-1(0)background than in wild type. Our results implicate these kinases as potential regulators of microRNA-induced silencing complex activity during postdauer development inC. elegans.