Mechanism Of Action And Functions Of Glis 1-3

Glis的作用机制和功能1-3

• 批准号: 7007508
• 负责人: Anton M Jetten
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7007508
• 关键词: biological signal transduction; bone morphogenetic proteins; developmental genetics; embryogenesis; fibroblast growth factor; gene expression; genetically modified animals; human tissue; laboratory mouse; nuclear receptors; protein structure function; transcription factor; yeast two hybrid system

Glis1-3 are novel genes recently identified in our laboratory. The Glis1-3 genes encode Kruppel-like zinc finger proteins containing five tandem zinc finger motifs that exhibit highest homology with those of members of the Gli and Zic subfamilies of Kruppel-like proteins. In addition, the zinc finger domain of Glis1 and -3 exhibit high homology with that of Drosophila gleeful/lame duck suggesting that it may be the Drosophila homologue of Glis1 and -3. Northern blot analysis showed that expression of the Glis1 and 2 mRNA is most abundant in adult kidney while Glis3 is expressed in several tissues. Whole mount in situ hybridization on mouse embryos demonstrated that Glis1-3 are expressed in a temporal and spatial manner during development. Glis1 expression was most prominent in several defined structures of mesodermal lineage, including craniofacial regions, branchial arches, somites, vibrissal and hair follicles, limb buds, and myotomes suggesting a role at different stages of development. Glis2 was expressed in kidney and neural tube suggesting a role in neurogenesis and kidney development.Glis3 is expressed in specific regions in developing kidney and testis and in a highly dynamic pattern during neurulation. From E11.5 through E12.5 Glis3 was strongly expressed in the interdigital regions, which are fated to undergo apoptosis. The temporal and spatial pattern of Glis1-3 expression observed during embryonic development suggests that they may play a critical role in the regulation of a variety of cellular processes during development.Confocal microscopic analysis showed that Glis1-3 are localized to the nucleus. The punctated pattern suggests that they are part of a larger nuclear protein complex. The zinc finger region in Glis plays an important role in the nuclear localization of these proteins. Electrophoretic mobility shift assays demonstrated that Glis1-3 are able to bind oligonucleotides containing the Gli-binding site consensus sequence GACCACCCAC. Although monohybrid analysis showed that in several cell types Glis1-3 are unable to induce transcription of a reporter, deletion mutant analysis revealed the presence of a strong activation and repressor functions suggesting that these proteins can function as repressors and activators of transcription. Glis3 was found to interact with GLi1 suggesting interaction between the Glis and Gli signaling pathways. Our results suggest that Glis1-3 may play a critical role in the control of gene expression during specific stages of embryonic development. Our hypothesis is that these proteins may act up- and/or downstream of sonic hedgehog, Wnt, BMP, or FGF signaling pathways. In addition to links between these signaling pathways, Glis proteins interact with the nuclear receptor RORgamma signaling pathway: Glis3 can suppress the transcriptional activation by RORgamma.

GLIS1-3是最近在我们的实验室中发现的新基因。 GLIS1-3基因编码含有五个串联锌指基序的Kruppel样锌指蛋白，它们与Kruppel样蛋白的GLI和ZIC亚家族成员表现出最高的同源性。此外，GLIS1和-3的锌指域与果蝇Gleeful/Lea Duck的同源性表现出很高的同源性，这表明它可能是Glis1和-3的果蝇同源物。北印迹分析表明，在成年肾脏中GLIS1和2 mRNA的表达最丰富，而GLIS3在几个组织中表达。小鼠胚胎上的整个原位杂交表明，GLIS1-3在发育过程中以时间和空间方式表达。 GLIS1表达在中胚层谱系的几个定义结构中最为突出，包括颅面区域，分支拱门，物种，振动和毛囊，肢体芽和肌肉群，表明在不同的发育阶段发挥作用。 GLIS2在肾脏和神经管中表达，表明在神经发生和肾脏发育中起作用。GLIS3在发育中的肾脏和睾丸以及在神经过程中具有高度动态的特定区域表达。从E11.5到E12.5 GLIS3在数字间区域强烈表达，这些区域被命中凋亡。在胚胎发育过程中观察到的GLIS1-3表达的时间和空间模式表明，它们可能在发育过程中各种细胞过程的调节中起关键作用。连接微观分析表明，Glis1-3定位于核。穿刺模式表明它们是较大的核蛋白质复合物的一部分。 GLI中的锌指区域在这些蛋白质的核定位中起重要作用。电泳迁移率转移分析表明，GLIS1-3能够结合含有Gli结合位点共识序列Gaccacccac的寡核苷酸。尽管单杂交分析表明，在几种细胞类型中，GLIS1-3无法诱导记者的转录，但缺失突变体分析表明存在强激活和阻遏物功能，表明这些蛋白质可以充当转录的阻遏物和激活剂。发现GLIS3与GLI1相互作用，表明GLI和GLI信号通路之间的相互作用。我们的结果表明，GLIS1-3在胚胎发育的特定阶段的控制基因表达中可能起关键作用。我们的假设是，这些蛋白质可能在Sonic Hedgehog，Wnt，BMP或FGF信号传导途径的向上和/或下游作用。 除了这些信号通路之间的联系外，Glis蛋白还与核受体Rorgamma信号通路相互作用：GLIS3可以抑制Rorgamma的转录激活。