Zhang Jun, Schulze Karen L, Hiesinger P Robin, Suyama Kaye, Wang Stream, Fish Matthew, Acar Melih, Hoskins Roger A, Bellen Hugo J, Scott Matthew P
Department of Developmental Biology, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, California 94305, USA.
Genetics. 2007 Jun;176(2):1307-22. doi: 10.1534/genetics.106.066761. Epub 2007 Apr 3.
Rab proteins are small GTPases that play important roles in transport of vesicle cargo and recruitment, association of motor and other proteins with vesicles, and docking and fusion of vesicles at defined locations. In vertebrates, >75 Rab genes have been identified, some of which have been intensively studied for their roles in endosome and synaptic vesicle trafficking. Recent studies of the functions of certain Rab proteins have revealed specific roles in mediating developmental signal transduction. We have begun a systematic genetic study of the 33 Rab genes in Drosophila. Most of the fly proteins are clearly related to specific vertebrate proteins. We report here the creation of a set of transgenic fly lines that allow spatially and temporally regulated expression of Drosophila Rab proteins. We generated fluorescent protein-tagged wild-type, dominant-negative, and constitutively active forms of 31 Drosophila Rab proteins. We describe Drosophila Rab expression patterns during embryogenesis, the subcellular localization of some Rab proteins, and comparisons of the localization of wild-type, dominant-negative, and constitutively active forms of selected Rab proteins. The high evolutionary conservation and low redundancy of Drosophila Rab proteins make these transgenic lines a useful tool kit for investigating Rab functions in vivo.
Rab蛋白是一类小GTP酶，在囊泡货物运输、马达蛋白及其他蛋白与囊泡的募集和结合，以及囊泡在特定位置的对接和融合过程中发挥重要作用。在脊椎动物中，已鉴定出超过75个Rab基因，其中一些因在内涵体和突触囊泡运输中的作用而得到深入研究。最近对某些Rab蛋白功能的研究揭示了它们在介导发育信号转导中的特定作用。我们已开始对果蝇中的33个Rab基因进行系统的遗传学研究。大多数果蝇蛋白与特定的脊椎动物蛋白明显相关。我们在此报告创建了一组转基因果蝇品系，可实现果蝇Rab蛋白在空间和时间上的调控表达。我们生成了31种果蝇Rab蛋白的荧光蛋白标记的野生型、显性负性和组成型激活形式。我们描述了胚胎发育过程中果蝇Rab的表达模式、一些Rab蛋白的亚细胞定位，以及所选Rab蛋白的野生型、显性负性和组成型激活形式的定位比较。果蝇Rab蛋白的高度进化保守性和低冗余性使这些转基因品系成为研究体内Rab功能的有用工具集。
