Lee Bruce P, Jones Bradley W
Molecular Neurobiology Program, Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, NY 10016, USA.
Mech Dev. 2005 Jun;122(6):849-62. doi: 10.1016/j.mod.2005.01.002. Epub 2005 Mar 11.
reversed polarity (repo) is a putative target gene of glial cells missing (gcm), the primary regulator of glial cell fate in Drosophila. Transient expression of Gcm is followed by maintained expression of repo. Multiple Gcm binding sites are found in repo upstream DNA. However, while repo is expressed in Gcm positive glia, it is not expressed in Gcm positive hemocytes. These observations suggest factors in addition to Gcm are required for repo expression. Here we have undertaken an analysis of the cis-regulatory DNA elements of repo using lacZ reporter activity in transgenic embryos. We have found that a 4.2 kb DNA region upstream of the repo start site drives the wild-type repo expression pattern. We show that expression is dependent on multiple Gcm binding sites. By ectopically expressing Repo, we show that Repo can regulate its own enhancer. Finally, by systematically analyzing fragments of repo upstream DNA, we show that expression is dependent on multiple elements that are responsible for activity in subsets of glia, as well as repressing inappropriate expression in the epidermis. Our results suggest that Gcm acts synergistically with other factors to control repo transcription in glial cells.
反向极性(repo)是果蝇中神经胶质细胞命运的主要调节因子——神经胶质细胞缺失(gcm)的一个假定靶基因。Gcm的瞬时表达之后是repo的持续表达。在repo上游DNA中发现了多个Gcm结合位点。然而,虽然repo在Gcm阳性神经胶质细胞中表达,但在Gcm阳性血细胞中不表达。这些观察结果表明,除了Gcm之外,repo表达还需要其他因子。在这里,我们利用转基因胚胎中的lacZ报告基因活性对repo的顺式调控DNA元件进行了分析。我们发现,repo起始位点上游4.2 kb的DNA区域驱动野生型repo的表达模式。我们表明,表达依赖于多个Gcm结合位点。通过异位表达Repo,我们表明Repo可以调节其自身的增强子。最后,通过系统分析repo上游DNA片段,我们表明表达依赖于多个元件,这些元件负责在神经胶质细胞亚群中的活性,并抑制表皮中的不适当表达。我们的结果表明,Gcm与其他因子协同作用,以控制神经胶质细胞中repo的转录。