The bld1 mutation identifies the Chlamydomonas osm-6 homolog as a gene required for flagellar assembly

AbstractInsertional mutagenesis procedures in Chlamydomonas[1] have facilitated the identification and characterization of dozens of genes required for the assembly and motility of flagella in Chlamydomonas[2–6]. 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[7], 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[8] whose product is necessary for the assembly of a set of sensory cilia [9]. The protein product of the BLD1 gene corresponds to IFT52 [10], 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) [11, 12]. The BLD1 RNA transcript is upregulated upon flagellar amputation, as observed for many other genes encoding flagellar proteins [13]. These results demonstrate that the function of the IFT52 protein in Chlamydomonas is essential for the assembly and/or maintenance of the flagella