Piao Tian, Luo Minna, Wang Liang, Guo Yan, Li De, Li Peng, Snell William J, Pan Junmin
Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China.
Proc Natl Acad Sci U S A. 2009 Mar 24;106(12):4713-8. doi: 10.1073/pnas.0808671106. Epub 2009 Mar 5.
Cilia and flagella are dynamic organelles that are assembled and disassembled during cell differentiation, during stress, and during the cell cycle. Although intraflagellar transport (IFT) is well documented to be responsible for transport of ciliary/flagellar precursors from the cell body to the flagella, little is known about the molecular mechanisms for mobilizing the cell body-localized precursors to make them available for transport during organelle assembly or for disassembling the microtubule-based axoneme during shortening. Here, we show that Chlamydomonas kinesin-13 (CrKinesin-13), a member of the kinesin-13 family of microtubule depolymerizing kinesins best known for their roles in the cell cycle, functions in flagellar disassembly and flagellar assembly. Activation of a cell to generate new flagella induces rapid phosphorylation of CrKinesin-13, and activation of flagellar shortening induces the immediate transport of CrKinesin-13 via intraflagellar transport from the cell body into the flagella. Cells depleted of CrKinesin-13 by RNAi assemble flagella after cell division but are incapable of the rapid assembly of flagella that normally occurs after flagellar detachment. Furthermore, they are inhibited in flagellar shortening. Thus, CrKinesin-13 is dynamically regulated during flagellar assembly and disassembly in Chlamydomonas and functions in each.
纤毛和鞭毛是动态细胞器,在细胞分化、应激和细胞周期过程中会进行组装和拆卸。尽管轴丝内运输(IFT)已被充分证明负责将纤毛/鞭毛前体从细胞体运输到鞭毛,但对于动员细胞体定位的前体以使其在细胞器组装期间可用于运输或在缩短过程中拆卸基于微管的轴丝的分子机制知之甚少。在这里,我们表明衣藻驱动蛋白-13(CrKinesin-13),一种驱动蛋白-13家族的微管解聚驱动蛋白,以其在细胞周期中的作用而闻名,在鞭毛拆卸和鞭毛组装中发挥作用。激活细胞以产生新的鞭毛会诱导CrKinesin-13的快速磷酸化,而鞭毛缩短的激活会诱导CrKinesin-13通过轴丝内运输从细胞体立即运输到鞭毛中。通过RNAi耗尽CrKinesin-13的细胞在细胞分裂后组装鞭毛,但无法快速组装通常在鞭毛脱离后发生的鞭毛。此外,它们在鞭毛缩短方面受到抑制。因此,CrKinesin-13在衣藻鞭毛组装和拆卸过程中受到动态调节,并在每个过程中发挥作用。