Functional expression cloning reveals proapoptotic role for protein phosphatase 4
Functional expression cloning reveals proapoptotic role for protein phosphatase 4
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DOI：
10.1038/sj.cdd.4401274
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发表时间：
2003-09-01
影响因子：
12.4
通讯作者：
Williams, GT
中科院分区：
文献类型：
--
作者：
Mourtada-Maarabouni, M;Kirkham, L;Williams, GT
功能表达克隆策略非常适合于分析细胞凋亡的分子控制。这种方法有两个关键优势。首先，它消除了有关蛋白质性质的先验假设，其次，它选择性地靶向与细胞凋亡控制有关的蛋白质，这些蛋白质影响细胞在生存和死亡之间的关键决定。将这种策略应用于分离赋予对地塞米松和γ-照射抗性的cDNA导致分离蛋白磷酸酶4（PP 4）的催化亚基的部分cDNA。在表达载体中用该部分cDNA转染的细胞下调PP 4，并且对地塞米松和UV辐射具有抗性，如通过膜完整性和集落形成测定所证明的。这些观察结果表明，PP 4在T淋巴细胞中起着重要的促凋亡作用。
Functional expression cloning strategies are highly suitable for the analysis of the molecular control of apoptosis. This approach has two critical advantages. Firstly, it eliminates prior assumptions about the properties of the proteins involved, and, secondly, it selectively targets proteins that are causally involved in apoptosis control and which affect the crucial cellular decision between survival and death. The application of this strategy to the isolation of cDNAs conferring resistance to dexamethasone and gamma-irradiation resulted in the isolation of a partial cDNA for the catalytic subunit of protein phosphatase 4 (PP4). Cells transfected with this partial cDNA in an expression vector downregulated PP4 and were resistant to both dexamethasone and UV radiation, as demonstrated by both membrane integrity and colony-forming assays. These observations suggest that PP4 plays an important proapoptotic role in T lymphocytes.