Tam Lai-Wa, Wilson Nedra F, Lefebvre Paul A
Department of Plant Biology, University of Minnesota, St. Paul, MN 55108, USA.
J Cell Biol. 2007 Mar 12;176(6):819-29. doi: 10.1083/jcb.200610022.
Little is known about how cells regulate the size of their organelles. In this study, we find that proper flagellar length control in Chlamydomonas reinhardtii requires the activity of a new member of the cyclin-dependent kinase (CDK) family, which is encoded by the LF2 (long flagella 2) gene. This novel CDK contains all of the important residues that are essential for kinase activity but lacks the cyclin-binding motif PSTAIRE. Analysis of genetic lesions in a series of lf2 mutant alleles and site-directed mutagenesis of LF2p reveals that improper flagellar length and defective flagellar assembly correlate with the extent of disruption of conserved kinase structures or residues by mutations. LF2p appears to interact with both LF1p and LF3p in the cytoplasm, as indicated by immunofluorescence localization, sucrose density gradients, cell fractionation, and yeast two-hybrid experiments. We propose that LF2p is the catalytic subunit of a regulatory kinase complex that controls flagellar length and flagellar assembly.
关于细胞如何调节其细胞器大小,人们所知甚少。在本研究中,我们发现莱茵衣藻中鞭毛长度的正常控制需要细胞周期蛋白依赖性激酶(CDK)家族一个新成员的活性,该成员由LF2(长鞭毛2)基因编码。这种新型CDK包含激酶活性所必需的所有重要残基,但缺乏细胞周期蛋白结合基序PSTAIRE。对一系列lf2突变等位基因中的遗传损伤分析以及LF2p的定点诱变表明,鞭毛长度异常和鞭毛组装缺陷与突变对保守激酶结构或残基的破坏程度相关。免疫荧光定位、蔗糖密度梯度、细胞分级分离和酵母双杂交实验表明,LF2p似乎在细胞质中与LF1p和LF3p相互作用。我们提出,LF2p是一种调节激酶复合物的催化亚基,该复合物控制鞭毛长度和鞭毛组装。