Drosophila genome-wide RNAi screen identifies multiple regulators of HIF-dependent transcription in hypoxia.
Drosophila genome-wide RNAi screen identifies multiple regulators of HIF-dependent transcription in hypoxia.
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DOI：
10.1371/journal.pgen.1000994
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发表时间：
2010-06-24
期刊：
影响因子：
4.5
通讯作者：
Wappner P
中科院分区：
文献类型：
--
作者：
Dekanty A;Romero NM;Bertolin AP;Thomas MG;Leishman CC;Perez-Perri JI;Boccaccio GL;Wappner P
低氧诱导因子（HIF）是一个进化上保守的α-β异二聚体转录因子家族，其诱导多种基因响应低氧张力。介导氧依赖性HIF调节的分子机制在α亚基水平上起作用，控制蛋白质稳定性、亚细胞定位和转录辅激活因子的募集。我们在果蝇细胞中进行了无偏见的全基因组RNA干扰（RNAi）筛选，旨在鉴定HIF活性所需的基因。经过3轮筛选，30个基因成为低氧中关键的HIF调节因子，其中大多数以前与HIF生物学无关。基因列表包括染色质重塑复合物、转录延伸因子和翻译调节因子的组分。一个引人注目的发现是argonaute 1（ago 1）基因，它是microRNA（miRNA）翻译沉默机制的核心元件。进一步的研究证实了miRNA机制在HIF依赖性转录中的生理作用。这项研究揭示了HIF调节的新机制的发生，这可能有助于开发治疗干预HIF相关病理（包括心脏病发作、癌症和中风）的新策略。细胞对低氧（缺氧）的适应是与重要疾病相关的生理反应，包括心脏病发作，中风，癌症和糖尿病。调节人类适应缺氧的机制与不同动物物种（包括小鼠、蠕虫和昆虫）的机制几乎相同。细胞对缺氧反应的主要调节因子是一种名为HIF的转录因子，它诱导一组基因介导对氧饥饿的适应。尽管已知HIF的调节主要发生在蛋白质降解和转录辅激活因子募集的水平，但以前没有进行HIF调节剂的全面筛选。在这项工作中，我们进行了基于RNAi的果蝇基因组筛选，寻找HIF活性所需的基因。在细胞培养系统中进行的这种筛选导致了HIF的30个关键调节因子的定义，其中大多数以前与缺氧生物学无关。筛选的命中物包括染色质重塑复合物、转录延伸因子和翻译调节因子的组分。我们的研究结果开启了对HIF调控进行详细研究的可能性，这可能导致重要人类疾病的新治疗策略。
Hypoxia-inducible factors (HIFs) are a family of evolutionary conserved alpha-beta heterodimeric transcription factors that induce a wide range of genes in response to low oxygen tension. Molecular mechanisms that mediate oxygen-dependent HIF regulation operate at the level of the alpha subunit, controlling protein stability, subcellular localization, and transcriptional coactivator recruitment. We have conducted an unbiased genome-wide RNA interference (RNAi) screen in Drosophila cells aimed to the identification of genes required for HIF activity. After 3 rounds of selection, 30 genes emerged as critical HIF regulators in hypoxia, most of which had not been previously associated with HIF biology. The list of genes includes components of chromatin remodeling complexes, transcription elongation factors, and translational regulators. One remarkable hit was the argonaute 1 (ago1) gene, a central element of the microRNA (miRNA) translational silencing machinery. Further studies confirmed the physiological role of the miRNA machinery in HIF–dependent transcription. This study reveals the occurrence of novel mechanisms of HIF regulation, which might contribute to developing novel strategies for therapeutic intervention of HIF–related pathologies, including heart attack, cancer, and stroke. Adaptation of cells to low oxygen (hypoxia) is a physiological response related to important diseases, including heart attacks, stroke, cancer, and diabetes. The mechanisms that mediate adaptation to hypoxia in humans are almost identical to those operating in diverse animal species, including mice, worms, and insects. The master regulator of cellular responses to hypoxia is a transcription factor named HIF, which induces a set of genes that mediate adaptation to oxygen starvation. Although it is known that regulation of HIF occurs mainly at the level of protein degradation and transcriptional coactivator recruitment, a comprehensive screen for HIF regulators has not been performed before. In this work, we have conducted an RNAi-based screen of the genome of the fruit fly Drosophila melanogaster, searching for genes that are required for HIF activity. This screen carried out in a cell culture system led to the definition of 30 critical regulators of HIF, most of which have not been associated with hypoxia biology before. The hits of the screen included components of chromatin remodeling complexes, transcription elongation factors, and translational regulators. Our results open the possibility of performing detailed studies on HIF regulation that may lead to novel therapeutic strategies for important human diseases.
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影响因子：
11.4
作者：
Bauer, A;Chauvet, S;Pradel, J
通讯作者：
Pradel, J
影响因子：
56.9
作者：
Jaakkola, P;Mole, DR;Ratcliffe, PJ
通讯作者：
Ratcliffe, PJ
影响因子：
10.5
作者：
Bemis, L;Chan, DA;Zundel, W
通讯作者：
Zundel, W
DOI：
10.1073/pnas.110149597
发表时间：
2000-06-06
影响因子：
11.1
作者：
Clemens, JC;Worby, CA;Dixon, JE
通讯作者：
Dixon, JE
影响因子：
64.8
作者：
Bernardi, Rosa;Guernah, Ilhem;Pandolfi, Pier Paolo
通讯作者：
Pandolfi, Pier Paolo