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转录因子 Krüppel 同源物 1 和 Hairy 在蚊虫 Aedes aegypti 中共同作用于保幼激素/灭幼脲耐受介导的基因抑制。

Synergistic action of the transcription factors Krüppel homolog 1 and Hairy in juvenile hormone/Methoprene-tolerant-mediated gene-repression in the mosquito Aedes aegypti.

机构信息

Department of Entomology and Institute of Integrative Biology, University of California, Riverside, California, United States of America.

Department of Biological Sciences, Birla Institute of Technology and Science Pilani, K. K. Birla Goa Campus, Goa, India.

出版信息

PLoS Genet. 2019 Oct 29;15(10):e1008443. doi: 10.1371/journal.pgen.1008443. eCollection 2019 Oct.

DOI:10.1371/journal.pgen.1008443
PMID:31661489
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6818763/
Abstract

Arthropod-specific juvenile hormones control numerous essential functions in development and reproduction. In the dengue-fever mosquito Aedes aegypti, in addition to its role in immature stages, juvenile hormone III (JH) governs post-eclosion (PE) development in adult females, a phase required for competence acquisition for blood feeding and subsequent egg maturation. During PE, JH through its receptor Methoprene-tolerant (Met) regulate the expression of many genes, causing either activation or repression. Met-mediated gene repression is indirect, requiring involvement of intermediate repressors. Hairy, which functions downstream of Met in the JH gene-repression hierarchy, is one such factor. Krüppel-homolog 1, a zinc-finger transcriptional factor, is directly regulated by Met and has been implicated in both activation and repression of JH-regulated genes. However, the interaction between Hairy and Kr-h1 in the JH-repression hierarchy is not well understood. Our RNAseq-based transcriptomic analysis of the Kr-h1-depleted mosquito fat body revealed that 92% of Kr-h1 repressed genes are also repressed by Met, supporting the existence of a hierarchy between Met and Kr-h1 as previously demonstrated in various insects. Notably, 130 genes are co-repressed by both Kr-h1 and Hairy, indicating regulatory complexity of the JH-mediated PE gene repression. A mosquito Kr-h1 binding site in genes co-regulated by this factor and Hairy was identified computationally. Moreover, this was validated using electrophoretic mobility shift assays. A complete phenocopy of the effect of Met RNAi depletion on target genes could only be observed after Kr-h1 and Hairy double RNAi knockdown, suggesting a synergistic action between these two factors in target gene repression. This was confirmed using a cell-culture-based luciferase reporter assay. Taken together, our results indicate that Hairy and Kr-h1 not only function as intermediate downstream factors, but also act together in a synergistic fashion in the JH/Met gene repression hierarchy.

摘要

节肢动物特异性的保幼激素控制着发育和生殖过程中的许多重要功能。在登革热蚊子埃及伊蚊中，除了在不成熟阶段的作用外，保幼激素 III (JH) 还控制着成年雌蚊出茧后 (PE) 的发育，这是获得吸血和随后卵成熟能力的一个阶段。在 PE 期间，JH 通过其受体 Methoprene-tolerant (Met) 调节许多基因的表达，导致基因的激活或抑制。Met 介导的基因抑制是间接的，需要中间抑制物的参与。Hairy 作为 JH 基因抑制级联中的 Met 下游因子，就是这样一个因素。Krüppel-homolog 1，一种锌指转录因子，直接受 Met 调控，已被牵连到 JH 调控基因的激活和抑制中。然而，在 JH 抑制级联中 Hairy 和 Kr-h1 之间的相互作用还不是很清楚。我们基于 RNAseq 的转录组分析表明，在 Kr-h1 耗尽的蚊子脂肪体中，92%的 Kr-h1 抑制基因也受到 Met 的抑制，这支持了 Met 和 Kr-h1 之间存在级联关系，就像在各种昆虫中已经证明的那样。值得注意的是，有 130 个基因同时受到 Kr-h1 和 Hairy 的共同抑制，这表明 JH 介导的 PE 基因抑制存在调控复杂性。通过计算识别出了在这两个因子和 Hairy 共同调控的基因中存在的一个蚊子 Kr-h1 结合位点。此外，这通过电泳迁移率变动分析得到了验证。只有在 Kr-h1 和 Hairy 双重 RNAi 敲低后，才能观察到 Met RNAi 耗尽对靶基因的影响的完全表型复制，这表明这两个因子在靶基因抑制中存在协同作用。这通过基于细胞培养的荧光素酶报告基因检测得到了证实。总的来说，我们的研究结果表明，Hairy 和 Kr-h1 不仅作为中间下游因子发挥作用，而且在 JH/Met 基因抑制级联中还协同作用。

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