Crack Donna, Secombe Julie, Coombe Michelle, Brumby Anthony, Saint Robert, Richardson Helena
Center for the Molecular Genetics of Development and Department of Molecular Biosciences, University of Adelaide, Adelaide, South Australia, 5005, Australia.
Dev Biol. 2002 Jan 1;241(1):157-71. doi: 10.1006/dbio.2001.0496.
The Drosophila cyclin E (DmcycE) gene gives rise to two transcripts encoding proteins that differ at their N termini, DmcycEII and DmcycEI. This study presents the first in vivo dissection of Cyclin E function. Ectopic expression studies using N- and C-terminal deletions of DmcycEI revealed that a region of 322 residues surrounding the cyclin box is sufficient to induce entry of G1-arrested larval eye imaginal disc cells into S phase. Ectopic expression of DmcycEI in the eye disc has been previously shown to drive anterior, but not posterior, G1-phase cells within the morphogenetic furrow (MF) into S phase. Significantly, ectopic expression of DmcycEII and N-terminal deletions of DmcycEI were able to drive all G1 cells within the morphogenetic furrow into S phase, while a C-terminal deletion of DmcycEI could not. The p21 homolog Dacapo was shown by yeast two-hybrid, coimmunolocalization, and in vivo functional studies not to be the mediator of the DmcycEI inhibition in posterior part of the MF. Taken together, these results reveal a novel zone within the posterior region of the MF where DmcycEI but not DmcycEII function is inhibited, and suggest that DmcycEII is a more potent inducer of S phase.
果蝇细胞周期蛋白E(DmcycE)基因产生两种转录本,编码在N端不同的蛋白质,即DmcycEII和DmcycEI。本研究首次对细胞周期蛋白E的功能进行了体内剖析。使用DmcycEI的N端和C端缺失进行的异位表达研究表明,细胞周期蛋白框周围322个残基的区域足以诱导G1期停滞的幼虫眼成虫盘细胞进入S期。先前已证明,在眼盘中异位表达DmcycEI可促使形态发生沟(MF)内的前部而非后部G1期细胞进入S期。值得注意的是,DmcycEII的异位表达和DmcycEI的N端缺失能够促使形态发生沟内的所有G1期细胞进入S期,而DmcycEI的C端缺失则不能。通过酵母双杂交、共免疫定位和体内功能研究表明,p21同源物Dacapo不是MF后部DmcycEI抑制作用的介导因子。综上所述,这些结果揭示了MF后部区域内一个新的区域,其中DmcycEI而非DmcycEII的功能受到抑制,并表明DmcycEII是一种更强效的S期诱导剂。