Cytogenetics Laboratory, Department of Zoology, Banaras Hindu University, Varanasi, India.
Genetics. 2010 Apr;184(4):927-45. doi: 10.1534/genetics.109.113696. Epub 2010 Jan 11.
Following earlier reports on modulation of poly(Q) toxicity in Drosophila by the developmentally active and stress-inducible noncoding hsromega gene, we investigated possible mediators of this modulation. RNAi-mediated downregulation of the large nuclear hsromega-n transcript, which organizes the nucleoplasmic omega speckles, suppressed the enhancement of poly(Q) toxicity brought about by reduced availability of the heterogeneous nuclear ribonucleoprotein (hnRNP) Hrb87F and of the transcriptional regulator, cAMP response element binding (CREB) binding protein (CBP). Levels of CBP RNA and protein were reciprocally affected by hsromega transcript levels in eye disc cells. Our data suggest that CBP and hnRNPs like Hrb57A and Hrb87F physically interact with each other. In addition, downregulation of hsromega transcripts partially rescued eye damage following compromised proteasome activity, while overexpression of hsromega and/or poly(Q) proteins disrupted the proteasomal activity. Rescue of poly(Q) toxicity by hsromega-RNAi required normal proteasomal function. We suggest that hsromega-RNAi suppresses poly(Q) toxicity by elevating cellular levels of CBP, by enhancing proteasome-mediated clearance of the pathogenic poly(Q) aggregates, and by inhibiting induced apoptosis. The direct and indirect interactions of the hsromega transcripts with a variety of regulatory proteins like hnRNPs, CBP, proteasome, Drosophila inhibitor of apoptosis protein 1 (DIAP1), etc., reinforce the view that the noncoding hsromega RNA functions as a "hub" in cellular networks to maintain homeostasis by coordinating the functional availability of crucial cellular regulatory proteins.
在先前关于发育活跃和应激诱导的非编码 hsromega 基因对果蝇中多聚(Q)毒性的调节的报告之后,我们研究了这种调节的可能介导物。RNAi 介导的大核 hsromega-n 转录本的下调,该转录本组织核质 ω 斑点,抑制了由于异质核核糖核蛋白(hnRNP)Hrb87F 和转录调节剂 cAMP 反应元件结合(CREB)结合蛋白(CBP)的可用性降低而引起的多聚(Q)毒性的增强。CBP RNA 和蛋白水平在眼盘细胞中受 hsromega 转录本水平的反向影响。我们的数据表明,CBP 和 hnRNPs 如 Hrb57A 和 Hrb87F 彼此物理相互作用。此外,hsromega 转录本的下调部分挽救了蛋白酶体活性受损后的眼睛损伤,而 hsromega 和/或多聚(Q)蛋白的过表达破坏了蛋白酶体活性。hsromega-RNAi 对多聚(Q)毒性的挽救需要正常的蛋白酶体功能。我们认为,hsromega-RNAi 通过提高细胞内 CBP 的水平,通过增强蛋白酶体介导的清除致病性多聚(Q)聚集体,以及通过抑制诱导的细胞凋亡来抑制多聚(Q)毒性。hsromega 转录本与各种调节蛋白(如 hnRNPs、CBP、蛋白酶体、果蝇凋亡抑制蛋白 1(DIAP1)等)的直接和间接相互作用,加强了非编码 hsromega RNA 作为“枢纽”的观点在细胞网络中通过协调关键细胞调节蛋白的功能可用性来维持体内平衡。