Comprehensive split TEV based protein-protein interaction screening reveals TAOK2 as a key modulator of Hippo signalling to limit growth

Raw data for the focused split TEV protein-protein interaction screen on the Hippo pathway. For each interaction pair (bait/prey), six replicates (R1-R6) for fold change (FC) and control (Ctrl) conditions are provided. For data processing, resulting firefly values were divided by Renilla values to yield firefly/Renilla ratios for each well. Next, interaction data for each bait/prey interaction pair was normalized by calculating fold change values over a plate control (provided here as R1-R6). To compute fold changes, the averaged firefly/Renilla ratio of a bait/prey pair (bait-X-NTEV-tcs-GV::prey-Y-CTEV) was divided by the corresponding averaged firefly/Renilla ratio of the bait/RASSF1-noSARAH negative control (i.e., bait-X-NTEV-tcs-GV::R1-noSARAH-CTEV interaction) that was present on the same plate. Log2-transformed fold changes values were ranked and values above 1 (corresponds to a fold change of 2 and more) were considered as high-confidence hits. For the statistical analysis, each of the six replicates of firefly/Renilla ratios of a given bait/prey interaction pair were divided by the mean of the control bait/RASSF1-noSARAH negative control, yielding six normalized replicates per interaction. To compute statistical significance for each bait/prey pair, adjusted P values for multiple comparisons were calculated using the false discovery rate (FDR) correction method.

针对河马信号通路（Hippo pathway）的定向分裂TEV（split TEV）蛋白质相互作用筛选原始数据集。针对每一组相互作用对（诱饵蛋白/猎物蛋白，bait/prey），均提供了6次生物学重复（R1至R6）的倍数变化（fold change, FC）及对照（control, Ctrl）条件数据。数据处理环节中，首先将每孔检测得到的萤火虫荧光素酶（firefly）信号值除以海肾荧光素酶（Renilla）信号值，得到每孔的萤火虫/海肾荧光素酶比值；随后，通过计算相对于板内对照的倍数变化值，对每一组诱饵/猎物蛋白相互作用对的交互数据进行标准化处理（本次数据集的板内对照为R1至R6）。在计算倍数变化时，将某一诱饵/猎物蛋白对（bait-X-NTEV-tcs-GV::prey-Y-CTEV）的平均萤火虫/海肾荧光素酶比值，除以同一板内存在的诱饵/RASSF1-noSARAH阴性对照（即bait-X-NTEV-tcs-GV::R1-noSARAH-CTEV相互作用）的平均萤火虫/海肾荧光素酶比值。对经log₂转换后的倍数变化值进行排序，其中数值大于1（对应倍数变化≥2）的结果被判定为高可信度阳性命中。统计分析环节中，将给定诱饵/猎物蛋白相互作用对的6次重复的萤火虫/海肾荧光素酶比值，分别除以对照诱饵/RASSF1-noSARAH阴性对照的平均值，最终得到每一组相互作用的6次标准化重复数据。为计算每一组诱饵/猎物蛋白对的统计学显著性，采用错误发现率（false discovery rate, FDR）校正方法计算多重比较校正后的P值。