Deane J A, Cole D G, Seeley E S, Diener D R, Rosenbaum J L
Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520, USA.
Curr Biol. 2001 Oct 16;11(20):1586-90. doi: 10.1016/s0960-9822(01)00484-5.
Intraflagellar transport (IFT) is a motility in which particles composed of at least 17 polypeptides move underneath the flagellar membrane. Anterograde (outward) and retrograde (inward) movements of these IFT particles are mediated by FLA10 kinesin-II and cytoplasmic dynein DHC1b, respectively. Mutations affecting IFT particle polypeptides or motors result in the inability to assemble flagella. IFT particles and the motors moving them are located principally around the basal bodies as well as in the flagella. Here, we clone the cDNA encoding one of the IFT particle proteins, IFT52, and show by immunofluorescence that while some IFT52 is in the flagella, the majority is found in two horseshoe-shaped rings around the basal bodies. Immunoelectron microscopy indicates that IFT52 is associated with the periphery of the transitional fibers, which extend from the distal portion of the basal body to the cell membrane and demarcate the entrance to the flagellar compartment. This localization suggests that the transitional fibers form a docking complex for the IFT particles destined for the flagellum. Finally, the flagellaless mutant bld1 completely lacks IFT52 due to a deletion in the gene encoding IFT52.
鞭毛内运输(IFT)是一种运动方式,由至少17种多肽组成的颗粒在鞭毛膜下移动。这些IFT颗粒的顺行(向外)和逆行(向内)运动分别由FLA10驱动蛋白-II和细胞质动力蛋白DHC1b介导。影响IFT颗粒多肽或马达的突变会导致无法组装鞭毛。IFT颗粒及其移动的马达主要位于基体周围以及鞭毛中。在这里,我们克隆了编码一种IFT颗粒蛋白IFT52的cDNA,并通过免疫荧光显示,虽然一些IFT52存在于鞭毛中,但大部分位于基体周围的两个马蹄形环中。免疫电子显微镜表明,IFT52与过渡纤维的外周相关联,过渡纤维从基体的远端延伸到细胞膜,并划定了鞭毛区室的入口。这种定位表明,过渡纤维为运往鞭毛的IFT颗粒形成了一个对接复合体。最后,无鞭毛突变体bld1由于编码IFT52的基因缺失而完全缺乏IFT52。