PF16 encodes a protein with armadillo repeats and localizes to a single microtubule of the central apparatus in Chlamydomonas flagella.
PF16 encodes a protein with armadillo repeats and localizes to a single microtubule of the central apparatus in Chlamydomonas flagella.
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DOI：
10.1083/jcb.132.3.359
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发表时间：
1996-02
期刊：
影响因子：
--
通讯作者：
Lefebvre PA
中科院分区：
文献类型：
--
作者：
Smith EF;Lefebvre PA
一些研究表明，中央微管对及其相关结构在调节鞭毛运动中起着重要作用。为了开始这些组件的分子分析，我们已经产生了中央装置缺陷型突变体在衣原体reinhardtii使用插入诱变。在我们的筛选中恢复的一个瘫痪突变体D2是先前鉴定的突变体pf 16的等位基因。突变的细胞有瘫痪的鞭毛，和中央器的C1微管是在孤立的轴丝失踪。我们已经克隆了野生型PF 16基因，并通过转化后拯救PF 16突变体来确认其身份。拯救的pf 16细胞在运动性和轴丝超微结构上是野生型的。获得PF 16的全长cDNA克隆并测序。使用预测的PF 16的566个氨基酸序列的数据库搜索表明，该蛋白质含有8个连续的犰狳重复序列。许多具有不同细胞功能的蛋白质也含有犰狳重复序列，包括pendulin，Rch 1，importin，SRP-1和犰狳。针对从克隆的cDNA表达的融合蛋白产生抗体。野生型鞭毛的免疫荧光标记表明，PF 16蛋白是本地化沿着鞭毛的长度，而免疫金标记进一步本地化PF 16蛋白到一个单一的微管的中央对。基于本地化的结果和该蛋白质中的犰狳重复序列的存在下，我们建议PF 16基因产物参与C1中央微管稳定性和鞭毛运动重要的蛋白质-蛋白质相互作用。
Several studies have indicated that the central pair of microtubules and their associated structures play a significant role in regulating flagellar motility. To begin a molecular analysis of these components we have generated central apparatus-defective mutants in Chlamydomonas reinhardtii using insertional mutagenesis. One paralyzed mutant recovered in our screen, D2, is an allele of a previously identified mutant, pf16. Mutant cells have paralyzed flagella, and the C1 microtubule of the central apparatus is missing in isolated axonemes. We have cloned the wild-type PF16 gene and confirmed its identity by rescuing pf16 mutants upon transformation. The rescued pf16 cells were wild-type in motility and in axonemal ultrastructure. A full-length cDNA clone for PF16 was obtained and sequenced. Database searches using the predicted 566 amino acid sequence of PF16 indicate that the protein contains eight contiguous armadillo repeats. A number of proteins with diverse cellular functions also contain armadillo repeats including pendulin, Rch1, importin, SRP-1, and armadillo. An antibody was raised against a fusion protein expressed from the cloned cDNA. Immunofluorescence labeling of wild-type flagella indicates that the PF16 protein is localized along the length of the flagella while immunogold labeling further localizes the PF16 protein to a single microtubule of the central pair. Based on the localization results and the presence of the armadillo repeats in this protein, we suggest that the PF16 gene product is involved in protein-protein interactions important for C1 central microtubule stability and flagellar motility.