生物谷报道:日前,芬兰赫尔辛基大学领导的一个研究小组克隆出人类90%以上的功能型蛋白激酶基因和一系列相应的缺乏催化活性的激酶,用来进行蛋白激酶的功能性研究。
研究人员人类基因组中分离克隆出蛋白激酶基因,该类基因通过磷酸化和去磷酸化调控其他蛋白,参与细胞信号传导过程。据估计,约1/4的激酶在人类癌症患病过程中发生重要作用。研究小组进一步利用他们克隆出的激酶基因组群进行了蛋白激酶的高通量筛选,获得了2个新的激酶。一个起到调节“刺猬”信号通路的作用,这一通路与多个类型的人类癌症密切相关;另一种新的激酶则与卡波氏肉瘤泡疹病毒的作用相关。
这项研究分离出的众多激酶基因将最终形成一个基因库,可为日后系统分析激酶信号在不同的细胞疾病模式中的作用奠定基础。而且,激酶同样可以成为药物治疗的靶标,通过了解激酶,人类可以进而对付各种各样的癌症。研究结果发表在2号出版的cell上面。(生物谷www.bioon.com)
生物谷推荐原始出处:
Cell, Vol 133, 537-548, 02 May 2008
Application of Active and Kinase-Deficient Kinome Collection for Identification of Kinases Regulating Hedgehog Signaling
Markku Varjosalo,1,2,3,8 Mikael Björklund,1,2,3,8 Fang Cheng,2 Heidi Syvänen,1,2,3 Teemu Kivioja,1,2,4 Sami Kilpinen,2,5 Zairen Sun,6 Olli Kallioniemi,2,5 Hendrik G. Stunnenberg,7 Wei-Wu He,6 Päivi Ojala,2, and Jussi Taipale1,2,3,
1 Department of Molecular Medicine, National Public Health Institute (KTL), FI00290 Helsinki, Finland
2 Genome-Scale Biology Program, Biomedicum Helsinki, Institute of Biomedicine, University of Helsinki, PO Box 63, FI-00014, Finland
3 High Throughput Center, University of Helsinki, PO Box 63, FI-00014, Finland
4 Department of Computer Science, University of Helsinki, PO Box 63, FI-00014, Finland
5 Medical Biotechnology Center, VTT Technical Research Centre of Finland and University of Turku, Itäinen Pitkäkatu 4A, FI-20520 Turku, Finland
6 OriGene Technologies Inc, Six Taft Court, Suite 100, Rockville, MD 20850, USA
7 Department of Molecular Biology, Radboud University Nijmegen, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands
Summary
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, GSK3β
. Our results establish kinome expression screening as a highly effective way to identify physiological signaling pathway components and genes involved in pathological signaling crosstalk.