Lee Kiho, Shim Jiwon, Lee Jihyun, Lee Junho
Department of Biological Sciences, Seoul National University, Seoul, South Korea 151-742.
G3 (Bethesda). 2013 Oct 3;3(10):1779-84. doi: 10.1534/g3.113.007898.
Although many critical roles of the RUNX family proteins have already been identified, little attention has been given to how these proteins interact with other factors. Elucidating RUNX protein interactions will help extend our understanding of their roles in normal development and tumorigenesis. In this study, we performed large-scale RNAi screening to identify genes that genetically interact with rnt-1, the sole homolog of RUNX protein in the nematode Caenorhabditis elegans. To this end, we took advantage of the fact that C. elegans can survive a severe loss of RNT-1 function with only mild phenotypes, and we looked for genes that caused a synthetic phenotype in the rnt-1 mutant background. We identified seven genes, three of which (cdk-8, cic-1, and sur-2) are involved in transcription, two of which (pgp-2 and cct-5) are involved in stress response, and two of which (D2045.7 and W09D10.4) are involved in signaling cascades, according to their functional gene ontology terms. We further confirmed that the CDK8-containing mediator complex genetically interacts with RNT-1 by showing that knockdown of each component of the CDK8 mediator complex caused a synthetic phenotype, that is, the exploded intestine through the vulva (Eiv) phenotype, in the rnt-1 mutant background. We also identified a putative target gene, acs-4, which is regulated by the RNT-1 and CDK8 mediator complex. Our results strengthen the notion that the CDK8 mediator complex may also act together with RUNX proteins in mammals.
尽管RUNX家族蛋白的许多关键作用已经得到确认,但这些蛋白如何与其他因子相互作用却很少受到关注。阐明RUNX蛋白的相互作用将有助于拓展我们对其在正常发育和肿瘤发生中作用的理解。在本研究中,我们进行了大规模RNA干扰筛选,以鉴定与线虫秀丽隐杆线虫中RUNX蛋白的唯一同源物rnt-1发生遗传相互作用的基因。为此,我们利用了秀丽隐杆线虫能够在RNT-1功能严重丧失的情况下仅表现出轻微表型而存活的这一事实,并寻找在rnt-1突变背景下导致合成表型的基因。根据其功能基因本体术语,我们鉴定出了七个基因,其中三个(cdk-8、cic-1和sur-2)参与转录,两个(pgp-2和cct-5)参与应激反应,两个(D2045.7和W09D10.4)参与信号级联反应。我们进一步证实,含CDK8的中介体复合物与RNT-1发生遗传相互作用,这是通过表明在rnt-1突变背景下,敲低CDK8中介体复合物的每个组分都会导致一种合成表型,即通过阴门的肠道爆裂(Eiv)表型来实现的。我们还鉴定出一个推定的靶基因acs-4,它受RNT-1和CDK8中介体复合物调控。我们的结果强化了这样一种观念,即CDK8中介体复合物在哺乳动物中可能也与RUNX蛋白共同发挥作用。