Ojani R, Fu X, Ahmed T, Liu P, Zhu J
Department of Biochemistry, Virginia Tech, Blacksburg, VA, USA.
Program of Genetics, Bioinformatics, and Computational Biology, Virginia Tech, Blacksburg, VA, USA.
Insect Mol Biol. 2018 Apr;27(2):268-278. doi: 10.1111/imb.12370. Epub 2018 Jan 4.
Krüppel homologue 1 (Kr-h1) is a zinc finger transcription factor that is upregulated in insects by juvenile hormone (JH) in metamorphosis and adult reproduction. The molecular function of Kr-h1 in reproduction remains largely unknown. Here we report that AaKr-h1 functions as an important transcription regulator in adult female Aedes aegypti mosquitoes. The amount of AaKr-h1 protein increases with rising JH levels after adult emergence, reaches its peak at 48 h after eclosion, then decreases gradually and disappears after blood feeding. RNA interference (RNAi)-mediated depletion of AaKr-h1 substantially reduced egg production after blood feeding. Using a chromatin immunoprecipitation cloning approach, we identified in vivo AaKr-h1 binding sites in previtellogenic female mosquitoes. Binding of AaKr-h1 to the target genes correlated with its protein abundance. Interestingly, RNAi experiments indicated that AaKr-h1 played distinct roles when it bound to individual target genes. For example, depletion of AaKr-h1 led to substantial upregulation of AAEL005545 and AAEL004444, but also significantly decreased the expression of AAEL005957 and AAEL013177 when compared with the control mosquitoes. In summary, AaKr-h1 directly binds to the regulatory regions of its target genes and acts as a transcriptional activator or a repressor in a promoter-specific manner.
克氏同源物1(Kr-h1)是一种锌指转录因子,在昆虫变态发育和成虫繁殖过程中受保幼激素(JH)上调。Kr-h1在繁殖中的分子功能仍 largely未知。在此,我们报道AaKr-h1在成年雌性埃及伊蚊中作为一种重要的转录调节因子发挥作用。成年羽化后,AaKr-h1蛋白的量随着JH水平的升高而增加,在羽化后48小时达到峰值,然后逐渐下降并在吸血后消失。RNA干扰(RNAi)介导的AaKr-h1缺失显著降低了吸血后的产卵量。使用染色质免疫沉淀克隆方法,我们在卵黄发生前的雌性蚊子体内鉴定出了AaKr-h1的结合位点。AaKr-h1与靶基因的结合与其蛋白丰度相关。有趣的是,RNAi实验表明,AaKr-h1与其个体靶基因结合时发挥不同的作用。例如,与对照蚊子相比,AaKr-h1的缺失导致AAEL005545和AAEL004444的显著上调,但也显著降低了AAEL005957和AAEL013177的表达。总之,AaKr-h1直接与其靶基因的调控区域结合,并以启动子特异性的方式作为转录激活因子或抑制因子发挥作用。