Brazelton W J, Amundsen C D, Silflow C D, Lefebvre P A
Department of Genetics, Cell, and Developmental Biology, 250 Bioscience Center, University of Minnesota, St. Paul, MN 55108, USA.
Curr Biol. 2001 Oct 16;11(20):1591-4. doi: 10.1016/s0960-9822(01)00485-7.
Insertional mutagenesis procedures in Chlamydomonas have facilitated the identification and characterization of dozens of genes required for the assembly and motility of flagella in Chlamydomonas. Many of these genes have been found to have homologs in animal systems. Here we describe a new gene required for flagellar assembly. Null mutants at the BLD1 locus assemble no flagella, and the flagellar membrane abuts the end of the transition zone distal to the basal body. Unlike mutants with basal body ultrastructural defects, such as bld2, bld1 mutants have normal basal bodies and cytoplasmic microtubule rootlets. The wild-type BLD1 gene was cloned by using DNA flanking the site of insertion of plasmid DNA in an insertional mutant; the cloned gene rescues the bld1 mutant phenotype upon transformation. The predicted BLD1 gene product is a 50.4 kDa protein with extensive regions of sequence similarity to the osm-6 gene of Caenorhabditis elegans whose product is necessary for the assembly of a set of sensory cilia. The protein product of the BLD1 gene corresponds to IFT52, a protein component of "raft" particles shown to undergo rapid transport up and down Chlamydomonas flagella between the flagellar membrane and the axoneme in a process known as intraflagellar transport (IFT). The BLD1 RNA transcript is upregulated upon flagellar amputation, as observed for many other genes encoding flagellar proteins. These results demonstrate that the function of the IFT52 protein in Chlamydomonas is essential for the assembly and/or maintenance of the flagella.
衣藻中的插入诱变程序有助于鉴定和表征衣藻鞭毛组装和运动所需的数十个基因。已发现其中许多基因在动物系统中有同源物。在此,我们描述了一个鞭毛组装所需的新基因。BLD1位点的无效突变体不组装鞭毛,并且鞭毛膜邻接基体远端的过渡区末端。与具有基体超微结构缺陷的突变体(如bld2)不同,bld1突变体具有正常的基体和细胞质微管小根。通过使用插入突变体中质粒DNA插入位点侧翼的DNA克隆了野生型BLD1基因;克隆的基因在转化后挽救了bld1突变体表型。预测的BLD1基因产物是一种50.4 kDa的蛋白质,与秀丽隐杆线虫的osm-6基因有广泛的序列相似区域,其产物对于一组感觉纤毛的组装是必需的。BLD1基因的蛋白质产物对应于IFT52,一种“筏”颗粒的蛋白质成分,已显示在称为鞭毛内运输(IFT)的过程中在鞭毛膜和轴丝之间在衣藻鞭毛中快速上下运输。如许多其他编码鞭毛蛋白的基因所观察到的,鞭毛切断后BLD1 RNA转录物上调。这些结果表明,衣藻中IFT52蛋白的功能对于鞭毛的组装和/或维持至关重要。