Tolley Nicholas, Sparkes Imogen A, Hunter Paul R, Craddock Christian P, Nuttall James, Roberts Lynne M, Hawes Chris, Pedrazzini Emanuela, Frigerio Lorenzo
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK.
Traffic. 2008 Jan;9(1):94-102. doi: 10.1111/j.1600-0854.2007.00670.x. Epub 2007 Dec 6.
We have cloned a member of the reticulon (RTN) family of Arabidopsis thaliana (RTNLB13). When fused to yellow fluorescent protein (YFP) and expressed in tobacco leaf epidermal cells, RTNLB13 is localized in the endoplasmic reticulum (ER). Coexpression of a soluble ER luminal marker reveals that YFP-tagged, myc-tagged or untagged RTNLB13 induces severe morphological changes to the lumen of the ER. We show, using fluorescence recovery after photobleaching (FRAP) analysis, that RTNLB13 overexpression greatly reduces diffusion of soluble proteins within the ER lumen, possibly by introducing constrictions into the membrane. In spite of this severe phenotype, Golgi shape, number and dynamics appear unperturbed and secretion of a reporter protein remains unaffected.
我们克隆了拟南芥网织蛋白(RTN)家族的一个成员(RTNLB13)。当与黄色荧光蛋白(YFP)融合并在烟草叶片表皮细胞中表达时,RTNLB13定位于内质网(ER)。一种可溶性内质网腔标记物的共表达表明,YFP标签、myc标签或无标签的RTNLB13会诱导内质网腔发生严重的形态变化。我们通过光漂白后荧光恢复(FRAP)分析表明,RTNLB13的过表达极大地降低了可溶性蛋白在内质网腔内的扩散,这可能是通过在膜上引入收缩来实现的。尽管有这种严重的表型,但高尔基体的形状、数量和动态似乎未受干扰,报告蛋白的分泌也未受影响。