Varjosalo Markku, Björklund Mikael, Cheng Fang, Syvänen Heidi, Kivioja Teemu, Kilpinen Sami, Sun Zairen, Kallioniemi Olli, Stunnenberg Hendrik G, He Wei-Wu, Ojala Päivi, Taipale Jussi
Department of Molecular Medicine, National Public Health Institute (KTL), FI00290 Helsinki, Finland.
Cell. 2008 May 2;133(3):537-48. doi: 10.1016/j.cell.2008.02.047.
To allow genome-scale identification of genes that regulate cellular signaling, we cloned >90% of all human full-length protein kinase cDNAs and constructed the corresponding kinase activity-deficient mutants. To establish the utility of this resource, we tested the effect of expression of the kinases on three different cellular signaling models. In all screens, many kinases had a modest but significant effect, apparently due to crosstalk between signaling pathways. However, the strongest effects were found with known regulators and novel components, such as MAP3K10 and DYRK2, which we identified in a mammalian Hedgehog (Hh) signaling screen. DYRK2 directly phosphorylated and induced the proteasome-dependent degradation of the key Hh pathway-regulated transcription factor, GLI2. MAP3K10, in turn, affected GLI2 indirectly by modulating the activity of DYRK2 and the known Hh pathway component, GSK3beta. Our results establish kinome expression screening as a highly effective way to identify physiological signaling pathway components and genes involved in pathological signaling crosstalk.
为了实现对调控细胞信号传导基因的全基因组规模鉴定，我们克隆了超过90%的人类全长蛋白激酶cDNA，并构建了相应的激酶活性缺陷型突变体。为了确定该资源的实用性，我们测试了激酶表达对三种不同细胞信号传导模型的影响。在所有筛选中，许多激酶都有适度但显著的影响，这显然是由于信号通路之间的串扰。然而，在已知调节因子和新成分中发现了最强的效应，例如我们在哺乳动物刺猬信号通路（Hh）筛选中鉴定出的MAP3K10和DYRK2。DYRK2直接磷酸化并诱导关键的Hh信号通路调节转录因子GLI2的蛋白酶体依赖性降解。反过来，MAP3K10通过调节DYRK2和已知的Hh信号通路成分GSK3β的活性间接影响GLI2。我们的结果表明激酶组表达筛选是鉴定生理信号通路成分和参与病理信号串扰基因的一种高效方法。
