Elam Candice A, Sale Winfield S, Wirschell Maureen
Department of Cell Biology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Methods Cell Biol. 2009;92:133-51. doi: 10.1016/S0091-679X(08)92009-4. Epub 2009 Nov 21.
The purpose of this chapter is to review the methodology and advances that have revealed conserved signaling proteins that are localized in the 9+2 ciliary axoneme for regulating motility. Diverse experimental systems have revealed that ciliary and eukaryotic flagellar motility is regulated by second messengers including calcium, pH, and cyclic nucleotides. In addition, recent advances in in vitro functional studies, taking advantage of isolated axonemes, pharmacological approaches, and biochemical analysis of axonemes have demonstrated that otherwise ubiquitous, conserved protein kinases and phosphatases are transported to and anchored in the axoneme. Here, we focus on the functional/pharmacological, genetic, and biochemical approaches in the model genetic system Chlamydomonas that have revealed highly conserved kinases, anchoring proteins (e.g., A-kinase anchoring proteins), and phosphatases that are physically located in the axoneme where they play a direct role in control of motility.
本章的目的是回顾那些揭示了定位于9+2纤毛轴丝中用于调节运动的保守信号蛋白的方法和进展。多种实验系统表明,纤毛和真核生物鞭毛的运动受包括钙、pH和环核苷酸在内的第二信使调节。此外,利用分离的轴丝、药理学方法以及轴丝的生化分析进行的体外功能研究的最新进展表明,那些在其他情况下普遍存在的保守蛋白激酶和磷酸酶被转运到轴丝并锚定在其中。在这里,我们重点关注模式遗传系统衣藻中的功能/药理学、遗传学和生化方法,这些方法揭示了高度保守的激酶、锚定蛋白(如A激酶锚定蛋白)和磷酸酶,它们位于轴丝中,在运动控制中发挥直接作用。