Multicilin drives centriole biogenesis via E2f proteins

Biochemistry suggests e2f4 forms a complex with the coiled-coiled protein multicilin (MCIDAS), a protein that is necessary and sufficient to specify multiciliated cells in vertebrates. Here, we performed RNAseq on Xenopus laevis ectoderm in the presence of multicilin alone or multicilin and a dominant-negative e2f4 construct. We also performed ChIPseq on e2f4 in the presence or absence of multicilin. Taken together, these data demonstrate how multicilin affects e2f4 genomic targets and their downstream transcription. RNAseq: misexpression of multicilin-HGR +/- dominant-negative e2f4 messenger RNAs in X. laevis animal caps, multicilin induced with dexamethasone at mid-stage 11 and harvested at 3 timepoints (3, 6, and 9 hours after induction, roughly corresponding to stages 13, 16, and 18) with 3 biological replicates. ChIPseq: misexpression of e2f4-GFP +/- multicilin-HGR messenger RNAs in X. laevis animal caps, multicilin induced at mid-stage 11 and harvested at one timepoint (6 hours after induction, roughly corresponding to stage 16), immunoprecipitated with anti-GFP and sequenced; 2 biological replicates. Background was input prior to IP.

生物化学研究显示，E2F4（e2f4）可与卷曲螺旋蛋白多纤毛细胞生成因子（multicilin，MCIDAS）形成复合物，该蛋白是脊椎动物中特化多纤毛细胞所必需且充分的调控因子。本研究针对单独表达多纤毛细胞生成因子，或共表达多纤毛细胞生成因子与显性负效E2F4（e2f4）构建体的非洲爪蟾（Xenopus laevis）外胚层样本开展了RNA测序（RNAseq）分析；同时在存在或缺失多纤毛细胞生成因子的条件下，针对E2F4（e2f4）进行了染色质免疫沉淀测序（ChIPseq）。综合上述实验数据，本数据集阐明了多纤毛细胞生成因子如何调控E2F4的基因组靶标及其下游转录过程。

RNA测序数据：在非洲爪蟾动物帽中异位表达多纤毛细胞生成因子-HGR（multicilin-HGR）±显性负效E2F4（e2f4）信使RNA（mRNA）；于胚胎发育11期中期用地塞米松诱导多纤毛细胞生成因子表达，分别在诱导后3、6、9小时（大致对应发育阶段13、16、18）三个时间点收集样本，设置3次生物学重复。

染色质免疫沉淀测序（ChIPseq）数据：在非洲爪蟾动物帽中异位表达E2F4-GFP（e2f4-GFP）±多纤毛细胞生成因子-HGR信使RNA（mRNA）；于胚胎发育11期中期诱导多纤毛细胞生成因子表达，在诱导后6小时（大致对应发育阶段16）收集样本，采用抗GFP抗体进行免疫沉淀后测序，设置2次生物学重复；免疫沉淀前的输入样本作为对照背景。