Loganathan Rajprasad, Lee Joslynn S, Wells Michael B, Grevengoed Elizabeth, Slattery Matthew, Andrew Deborah J
Department of Cell Biology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, United States.
Department of Biomedical Sciences, University of Minnesota Medical School, Duluth, MN 55812, United States.
Dev Biol. 2016 Jan 1;409(1):234-250. doi: 10.1016/j.ydbio.2015.10.016. Epub 2015 Oct 19.
Transcription factors affect spatiotemporal patterns of gene expression often regulating multiple aspects of tissue morphogenesis, including cell-type specification, cell proliferation, cell death, cell polarity, cell shape, cell arrangement and cell migration. In this work, we describe a distinct role for Ribbon (Rib) in controlling cell shape/volume increases during elongation of the Drosophila salivary gland (SG). Notably, the morphogenetic changes in rib mutants occurred without effects on general SG cell attributes such as specification, proliferation and apoptosis. Moreover, the changes in cell shape/volume in rib mutants occurred without compromising epithelial-specific morphological attributes such as apicobasal polarity and junctional integrity. To identify the genes regulated by Rib, we performed ChIP-seq analysis in embryos driving expression of GFP-tagged Rib specifically in the SGs. To learn if the Rib binding sites identified in the ChIP-seq analysis were linked to changes in gene expression, we performed microarray analysis comparing RNA samples from age-matched wild-type and rib null embryos. From the superposed ChIP-seq and microarray gene expression data, we identified 60 genomic sites bound by Rib likely to regulate SG-specific gene expression. We confirmed several of the identified Rib targets by qRT-pCR and/or in situ hybridization. Our results indicate that Rib regulates cell growth and tissue shape in the Drosophila salivary gland via a diverse array of targets through both transcriptional activation and repression. Furthermore, our results suggest that autoregulation of rib expression may be a key component of the SG morphogenetic gene network.
转录因子影响基因表达的时空模式,常常调控组织形态发生的多个方面,包括细胞类型特化、细胞增殖、细胞死亡、细胞极性、细胞形状、细胞排列和细胞迁移。在这项研究中,我们描述了带状蛋白(Rib)在果蝇唾液腺(SG)伸长过程中控制细胞形状/体积增加方面的独特作用。值得注意的是,rib突变体中的形态发生变化并未影响SG细胞的一般属性,如特化、增殖和凋亡。此外,rib突变体中细胞形状/体积的变化并未损害上皮特异性形态属性,如顶基极性和连接完整性。为了鉴定受Rib调控的基因,我们在特异性驱动SG中表达绿色荧光蛋白标记的Rib的胚胎中进行了染色质免疫沉淀测序(ChIP-seq)分析。为了了解ChIP-seq分析中鉴定出的Rib结合位点是否与基因表达变化相关,我们进行了微阵列分析,比较年龄匹配的野生型和rib基因缺失胚胎的RNA样本。通过叠加的ChIP-seq和微阵列基因表达数据,我们鉴定出60个被Rib结合的基因组位点,可能调控SG特异性基因表达。我们通过定量逆转录聚合酶链反应(qRT-pCR)和/或原位杂交证实了几个鉴定出的Rib靶标。我们的结果表明,Rib通过转录激活和抑制多种靶标来调控果蝇唾液腺中的细胞生长和组织形状。此外,我们的结果表明,rib表达的自我调节可能是SG形态发生基因网络的关键组成部分。