Yang Haiwang, Basquin Denis, Pauli Daniel, Oliver Brian
National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 50 South Drive, Bethesda, MD, 20892, USA.
Department of Genetics & Evolution, Sciences III, University of Geneva, Boulevard d'Yvoy 4, CH 1205, Geneva, Switzerland.
BMC Genomics. 2017 May 18;18(1):384. doi: 10.1186/s12864-017-3755-x.
Transcriptional elongation is a generic function, but is also regulated to allow rapid transcription responses. Following relatively long initiation and promoter clearance, RNA polymerase II can pause and then rapidly elongate following recruitment of positive elongation factors. Multiple elongation complexes exist, but the role of specific components in adult Drosophila is underexplored.
We conducted RNA-seq experiments to analyze the effect of RNAi knockdown of Suppressor of Triplolethal and lilliputian. We similarly analyzed the effect of expressing a dominant negative Cyclin-dependent kinase 9 allele. We observed that almost half of the genes expressed in adults showed reduced expression, supporting a broad role for the three tested genes in steady-state transcript abundance. Expression profiles following lilliputian and Suppressor of Triplolethal RNAi were nearly identical raising the possibility that they are obligatory co-factors. Genes showing reduced expression due to these RNAi treatments were short and enriched for genes encoding metabolic or enzymatic functions. The dominant-negative Cyclin-dependent kinase 9 profiles showed both overlapping and specific differential expression, suggesting involvement in multiple complexes. We also observed hundreds of genes with sex-biased differential expression following treatment.
Transcriptional profiles suggest that Lilliputian and Suppressor of Triplolethal are obligatory cofactors in the adult and that they can also function with Cyclin-dependent kinase 9 at a subset of loci. Our results suggest that transcriptional elongation control is especially important for rapidly expressed genes to support digestion and metabolism, many of which have sex-biased function.
转录延伸是一种普遍功能,但也受到调控以实现快速的转录反应。在经历相对较长的起始和启动子清除过程后,RNA聚合酶II可以暂停,然后在招募正性延伸因子后迅速延伸。存在多种延伸复合物,但特定成分在成年果蝇中的作用尚未得到充分研究。
我们进行了RNA测序实验,以分析RNA干扰敲低三隐性抑制因子和小人国蛋白的效果。我们同样分析了表达显性负性细胞周期蛋白依赖性激酶9等位基因的效果。我们观察到,成年果蝇中表达的几乎一半基因的表达量降低,这支持了所测试的三个基因在稳态转录本丰度中具有广泛作用。小人国蛋白和三隐性抑制因子RNA干扰后的表达谱几乎相同,这增加了它们是必需辅助因子的可能性。由于这些RNA干扰处理而表达量降低的基因较短,并且富含编码代谢或酶功能的基因。显性负性细胞周期蛋白依赖性激酶9的表达谱显示出重叠和特定的差异表达,表明其参与多种复合物。我们还观察到处理后数百个基因具有性别偏向的差异表达。
转录谱表明,小人国蛋白和三隐性抑制因子在成体中是必需的辅助因子,并且它们也可以在一部分基因座上与细胞周期蛋白依赖性激酶9一起发挥作用。我们的结果表明,转录延伸控制对于快速表达的基因以支持消化和代谢尤为重要,其中许多基因具有性别偏向的功能。