Functional characterization of PAS and HES family bHLH transcription factors during the metamorphosis of the red flour beetle, Tribolium castaneum.
Functional characterization of PAS and HES family bHLH transcription factors during the metamorphosis of the red flour beetle, Tribolium castaneum.
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DOI：
10.1016/j.gene.2009.08.003
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发表时间：
2009-12-01
期刊：
影响因子：
3.5
通讯作者：
Palli, Subba R.
中科院分区：
文献类型：
--
作者：
Bitra, Kavita;Tan, Anjiang;Dowling, Ashley;Palli, Subba R.
碱性螺旋-环-螺旋转录因子存在于动物、植物和真菌中，在细胞增殖、组织分化、发育和解毒的控制中起重要作用。虽然昆虫基因组中含有50多种螺旋环抑制转录因子，但只有少数几种的功能是已知的。RNAi技术已成为一种广泛使用的敲除基因表达以分析基因功能的工具。由于RNAi在赤拟谷盗中的作用良好，因此我们利用赤拟谷盗和RNAi技术研究了属于PAS和HES家族的19个bHLH转录因子在赤拟谷盗幼虫期的功能。栗色。我们搜索了T.鉴定了53个bHLH基因。系统发育分析将这53个基因分为PAS、HES、Myc/USF、Hand、Mesp、Shout、p48、NeuroD/Neurogenin、Atonal和AS-C等10个家族。在RNAi研究中，敲低PAS和HES家族的7个成员的表达影响了T.栗色。不能生长到达到临界重量以经历变态、不能完成幼虫-蛹或蛹-成虫蜕皮和异常翅发育是在RNAi昆虫中观察到的最常见的表型。在研究的bHLH转录因子中，类固醇受体辅激活因子（SRC）表现出最严重的表型。敲低编码SRC的基因的表达通过影响脂质代谢的调节而引起生长停滞。这些研究证明了RNAi在该模式昆虫中用于多基因家族成员的功能表征的能力。
The basic helix-loop-helix transcription factors are present in animals, plants and fungi and play important roles in the control of cellular proliferation, tissue differentiation, development and detoxification. Although insect genomes contain more than 50 Helix-Loop-Helix transcription factors, the functions of only a few are known. RNAi has become a widely used tool to knockdown the expression to analyze the function of genes. As RNAi works well in Tribolium castaneum, we utilized this insect and RNAi to determine functions of 19 bHLH transcription factors belonging to PAS and HES families during the larval stages of the red flour beetle, T. castaneum. We searched the genome sequence of T. castaneum and identified 53 bHLH genes. Phylogenetic analyses classified these 53 genes into ten families; PAS, HES, Myc/USF, Hand, Mesp, Shout, p48, NeuroD/Neurogenin, Atonal and AS-C. In RNAi studies, knocking-down the expression of seven members of the PAS and HES families affected the growth and development of T. castaneum. An inability to grow to reach critical weight to undergo metamorphosis, failure to complete larval-pupal or pupal-adult ecdysis and abnormal wing development are among the most common phenotypes observed in RNAi insects. Among the bHLH transcription factors studied, the steroid receptor coactivator (SRC) showed the most severe phenotypes. Knock-down in the expression of the gene coding for SRC caused growth arrest by affecting the regulation of lipid metabolism. These studies demonstrate the power of RNAi for functional characterization of members of the multigene families in this model insect.
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