Kawamori Akihito, Yamaguchi Masamitsu
Department of Applied Biology and Insect Biomedical Research Center, Kyoto Institute of Technology, Kyoto, Japan.
Cell Struct Funct. 2011;36(1):103-19. doi: 10.1247/csf.11004. Epub 2011 Apr 9.
DREF (DNA replication-related element-binding factor) plays important roles in replication and proliferation in vivo by regulating transcription of various genes. However, due to a lack of appropriate cell biological studies in vivo, roles of DREF during a single cell development are poorly understood. To address this question, we focused our attention on macrochaetes bristle development system. Utilizing cell lineage analysis focusing on a single posterior scutellar (PSC) macrochaete sensory organ precursor (SOP) lineages in combination with GAL4/UAS targeted expression system for DREF double strand RNA, we revealed that DREF plays no apparent role in differentiation process during SOP formation. Rather, DREF regulates the timing of asymmetric cell division but perhaps plays no direct role in differentiation during asymmetric cell division. Most importantly, DREF affected replication and growth in shaft cells and/or socket cells. Further analysis revealed that DREF is necessary but not sufficient for nuclear growth and protein synthesis in shaft cells. Finally, it could be demonstrated that DREF plays a critical role in regulating pcna transcription in endocycling shaft cells. All these results provide evidence that DREF plays critical roles, especially in endoreplication process of bristle development, at least in part by regulating the pcna gene expression.
DNA复制相关元件结合因子(DREF)通过调控多种基因的转录,在体内的复制和增殖过程中发挥重要作用。然而,由于缺乏合适的体内细胞生物学研究,DREF在单细胞发育过程中的作用仍知之甚少。为了解决这个问题,我们将注意力集中在大刚毛的发育系统上。利用针对单个后胸盾片(PSC)大刚毛感觉器官前体(SOP)谱系的细胞谱系分析,结合用于DREF双链RNA的GAL4/UAS靶向表达系统,我们发现DREF在SOP形成过程中的分化过程中没有明显作用。相反,DREF调节不对称细胞分裂的时间,但可能在不对称细胞分裂过程中的分化中不发挥直接作用。最重要的是,DREF影响毛干细胞和/或毛窝细胞的复制和生长。进一步分析表明,DREF对于毛干细胞的核生长和蛋白质合成是必要的,但并不充分。最后,可以证明DREF在调控内循环毛干细胞中的增殖细胞核抗原(pcna)转录中起关键作用。所有这些结果都提供了证据,表明DREF发挥关键作用,特别是在刚毛发育的内复制过程中,至少部分是通过调控pcna基因表达来实现的。