Global Analysis of Translational Reprogramming during Flagellar Regeneration Reveals the Role of Sphingolipid Metabolism in Ciliogenesis and Ciliopathies. Global Analysis of Translational Reprogramming during Flagellar Regeneration Reveals the Role of Sphingolipid Metabolism in Ciliogenesis and Ciliopathies

Protein synthesis is essential for cilium assembly, but little is known about translational reprogramming that occurs during ciliogenesis. Using ribosome footprinting, we perform global translatome profiling on Chlamydomonas during flagellar regeneration. Overall design: Chlamydomonas reinhardtii (strain CC-125) that were grown for 15, 30, 60, and 90 min after deflagellation to generate libraries for ribosome footprint sequencing (Ribo-seq) and RNA sequencing (RNA-seq) . A reference sample was collected from the same culture before deflagellation (Non-Treated). We also performed RNA sequencing for C.elegans mutants : sptl-1/sptl-2/sptl-3. Each strain was analyzed in biological duplicate.

蛋白质合成对于纤毛组装至关重要，但目前学界对纤毛发生过程中的翻译重编程仍缺乏深入了解。本研究采用核糖体足迹实验（ribosome footprinting），对鞭毛再生阶段的衣藻开展全局翻译组谱分析。
整体实验设计：将莱茵衣藻（Chlamydomonas reinhardtii，菌株CC-125）在脱鞭毛后分别培养15、30、60和90分钟，以此构建核糖体印迹测序（ribosome footprint sequencing, Ribo-seq）与RNA测序（RNA-seq）的测序文库。同时于脱鞭毛前从同一培养体系中采集对照样本（未处理组）。此外，本研究还对秀丽隐杆线虫（C. elegans）的sptl-1/sptl-2/sptl-3突变体开展了RNA测序。所有菌株均设置两次生物学重复。