Rupp Gerald, Porter Mary E
Department of Genetics, Cell Biology, and Development, University of Minnesota Medical School, 6-160 Jackson Hall, 321 Church Street SE, Minneapolis, MN 55455, USA.
J Cell Biol. 2003 Jul 7;162(1):47-57. doi: 10.1083/jcb.200303019.
The dynein regulatory complex (DRC) is an important intermediate in the pathway that regulates flagellar motility. To identify subunits of the DRC, we characterized a Chlamydomonas motility mutant obtained by insertional mutagenesis. The pf2-4 mutant displays an altered waveform that results in slow swimming cells. EM analysis reveals defects in DRC structure that can be rescued by reintroduction of the wild-type PF2 gene. Immunolocalization studies show that the PF2 protein is distributed along the length of the axoneme, where it is part of a discrete complex of polypeptides. PF2 is a coiled-coil protein that shares significant homology with a mammalian growth arrest-specific gene product (Gas11/Gas8) and a trypanosome protein known as trypanin. PF2 and its homologues appear to be universal components of motile axonemes that are required for DRC assembly and the regulation of flagellar motility. The expression of Gas8/Gas11 transcripts in a wide range of tissues may also indicate a potential role for PF2-related proteins in other microtubule-based structures.
动力蛋白调节复合体(DRC)是调节鞭毛运动途径中的一个重要中间体。为了鉴定DRC的亚基,我们对通过插入诱变获得的衣藻运动突变体进行了表征。pf2-4突变体表现出波形改变,导致细胞游动缓慢。电子显微镜分析揭示了DRC结构中的缺陷,通过重新引入野生型PF2基因可以挽救这些缺陷。免疫定位研究表明,PF2蛋白沿轴丝长度分布,它是一种离散多肽复合体的一部分。PF2是一种卷曲螺旋蛋白,与哺乳动物生长停滞特异性基因产物(Gas11/Gas8)和一种称为锥虫蛋白的锥虫蛋白具有显著同源性。PF2及其同源物似乎是运动轴丝的普遍组成部分,是DRC组装和鞭毛运动调节所必需的。Gas8/Gas11转录本在多种组织中的表达也可能表明PF2相关蛋白在其他基于微管的结构中具有潜在作用。