Department of Molecular Biology and Biophysics, University of Connecticut Health Center, Farmington, CT 06030-3305.
Mol Biol Cell. 2022 Jun 1;33(7):br10. doi: 10.1091/mbc.E22-02-0033. Epub 2022 Apr 7.
transitions from mitotically dividing vegetative cells to sexually competent gametes of two distinct mating types following nutrient deprivation. Gametes of opposite mating type interact via their cilia, initiating an intraciliary signaling cascade and ultimately fuse forming diploid zygotes. The process of gametogenesis is genetically encode, and a previous study revealed numerous significant changes in mRNA abundance during this life-cycle transition. Here we describe a proteomic analysis of cilia derived from vegetative and gametic cells of both mating types in an effort to assess the global changes that occur within the organelle during this process. We identify numerous membrane- and/or matrix-associated proteins in gametic cilia that were not detected in cilia from vegetative cells. This includes the pro-protein from which the GATI-amide gametic chemotactic modulator derives, as well as receptors, a dynamin-related protein, ammonium transporters, two proteins potentially involved in the intraciliary signaling cascade-driven increase in cAMP, and multiple proteins with a variety of interaction domains. These changes in ciliary composition likely directly affect the functional properties of this organelle as the cell transitions between life-cycle stages.
在营养缺乏的情况下,从有丝分裂分裂的营养细胞向两种不同交配型的有性配子转变。相反交配型的配子通过纤毛相互作用,启动纤毛内信号级联反应,最终融合形成二倍体合子。配子发生过程是遗传编码的,先前的研究表明,在这个生命周期转变过程中,mRNA 丰度发生了许多显著变化。在这里,我们描述了从两种交配型的营养细胞和配子细胞中衍生的纤毛的蛋白质组分析,以评估该过程中细胞器内发生的全局变化。我们在配子纤毛中鉴定到许多膜和/或基质相关蛋白,而在营养细胞的纤毛中则没有检测到这些蛋白。这包括从 GATI-酰胺配子趋化调节剂衍生的前蛋白,以及受体、一种与动力蛋白相关的蛋白、铵转运体、两种可能参与纤毛内信号级联驱动 cAMP 增加的蛋白,以及多种具有多种相互作用域的蛋白。这些纤毛组成的变化可能直接影响这个细胞器的功能特性,因为细胞在生命周期阶段之间转变。