Yoshioka Yasuhide, Nguyen Trong Tue, Fujiwara Shunsuke, Matsuda Risa, Valadez-Graham Viviana, Zurita Mario, Yamaguchi Masamitsu
Department of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto, Japan.
Genesis. 2012 Aug;50(8):599-611. doi: 10.1002/dvg.22017. Epub 2012 Mar 12.
The Drosophila Jun N-terminal kinase (JNK) gene basket (bsk) promoter contains a DNA replication-related element (DRE)-like sequence, raising the possibility of regulation by the DNA replication-related element-binding factor (DREF). Chromatin immunoprecipitation assays with anti-DREF IgG showed the bsk gene promoter region to be effectively amplified. Luciferase transient expression assays revealed the DRE-like sequence to be important for bsk gene promoter activity, and knockdown of DREF decreased the bsk mRNA level and the bsk gene promoter activity. Furthermore, knockdown of DREF in the notum compartment of wing discs by pannier-GAL4 and UAS-DREFIR resulted in a split thorax phenotype. Monitoring of JNK activity in the wing disc by LacZ expression in a puckered (puc)-LacZ enhancer trap line revealed the reduction in DREF knockdown clones. These findings indicate that DREF is involved in regulation of Drosophila thorax development via actions on the JNK pathway.
果蝇JUN氨基末端激酶（JNK）基因basket（bsk）启动子包含一个类DNA复制相关元件（DRE）序列，这增加了受DNA复制相关元件结合因子（DREF）调控的可能性。用抗DREF IgG进行的染色质免疫沉淀试验表明，bsk基因启动子区域能被有效扩增。荧光素酶瞬时表达试验显示，类DRE序列对bsk基因启动子活性很重要，敲低DREF会降低bsk mRNA水平和bsk基因启动子活性。此外，通过pannier - GAL4和UAS - DREFIR在翅盘的背板区域敲低DREF会导致胸部裂开的表型。通过在褶皱（puc）- LacZ增强子陷阱系中利用LacZ表达监测翅盘中的JNK活性，发现DREF敲低克隆中的活性降低。这些发现表明，DREF通过作用于JNK途径参与果蝇胸部发育的调控。
