Deconstructing Survivin: comprehensive genetic analysis of Survivin function by conditional knockout in a vertebrate cell line.
Deconstructing Survivin: comprehensive genetic analysis of Survivin function by conditional knockout in a vertebrate cell line.
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DOI：
10.1083/jcb.200806118
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发表时间：
2008-10-20
影响因子：
7.8
通讯作者：
Earnshaw, William C.
中科院分区：
文献类型：
--
作者：
Yue, Zuojun;Carvalho, Ana;Xu, Zhenjie;Yuan, Xuemei;Cardinale, Stefano;Ribeiro, Susana;Lai, Fan;Ogawa, Hiromi;Gudmundsdottir, Elisabet;Gassmann, Reto;Morrison, Ciaran G.;Ruchaud, Sandrine;Earnshaw, William C.
Survivin是一种关键的细胞蛋白，被认为在细胞凋亡调控、有丝分裂进程中发挥作用，或者两者兼而有之。在这项研究中，我们描述了在鸡DT40细胞中分离到两个Survivin基因的条件性敲除。缺乏Survivin的DT40细胞在胞质分裂失败后死于间期。然而，这些细胞对化疗药物依托泊苷表现出正常的敏感性。在零背景下表达Survivin突变体以重新评估几个关键残基的作用表明，如果DT40细胞唯一的Survivin缺失一个或多个被广泛研究的周期蛋白依赖的激酶磷酸化位点，那么它们就可以正常生长。据报道，一个或多个位点是与Smac或aurora B结合所必需的。核输出序列或二聚界面的突变使细胞对生长温度敏感。作为通过瞬时转染法研究蛋白质功能的其他研究的重要警告，三个Survivin突变体在野生型蛋白质存在的情况下无法定位，但在没有野生型蛋白质的情况下确实定位并确实支持生命。
Survivin is a key cellular protein thought to function in apoptotic regulation, mitotic progression, or possibly both. In this study, we describe the isolation of two conditional knockouts of the survivin gene in chicken DT40 cells. DT40 cells lacking Survivin die in interphase after failing to complete cytokinesis. However, these cells show normal sensitivity to the chemotherapeutic agent etoposide. Expression of Survivin mutants against a null background to reassess the role of several key residues reveals that DT40 cells can grow normally if their sole Survivin is missing a widely studied cyclin-dependent kinase phosphorylation site or sites reportedly essential for binding to Smac or aurora B. Mutations in the nuclear export sequence or dimerization interface render cells temperature sensitive for growth. As an important caveat for other studies in which protein function is studied by transient transfection, three of the Survivin mutants fail to localize in the presence of the wild-type protein but do localize and indeed support life in its absence.
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DOI：
10.1083/jcb.105.5.2053
发表时间：
1987-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Cooke CA;Heck MM;Earnshaw WC
通讯作者：
Earnshaw WC
DOI：
10.1083/jcb.153.4.865
发表时间：
2001-05-14
期刊：
The Journal of cell biology
影响因子：
--
作者：
Adams RR;Maiato H;Earnshaw WC;Carmena M
通讯作者：
Carmena M
DOI：
10.1073/pnas.0701140104
发表时间：
2007-03-13
影响因子：
11.1
作者：
Burkard, Mark E.;Randall, Catherine L.;Jallepalli, Prasad V.
通讯作者：
Jallepalli, Prasad V.
影响因子：
9.2
作者：
Fraser, AG;James, C;Hengartner, MO
通讯作者：
Hengartner, MO
影响因子：
7.5
作者：
Altieri, Dario C.
通讯作者：
Altieri, Dario C.