A large scale shRNA barcode screen identifies the circadian clock component ARNTL as putative regulator of the p53 tumor suppressor pathway.
A large scale shRNA barcode screen identifies the circadian clock component ARNTL as putative regulator of the p53 tumor suppressor pathway.
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DOI：
10.1371/journal.pone.0004798
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发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
Beijersbergen RL
中科院分区：
文献类型：
--
作者：
Mullenders J;Fabius AW;Madiredjo M;Bernards R;Beijersbergen RL
p53 肿瘤抑制基因在大约一半的人类癌症中发生突变，但 p53 通路被认为在绝大多数癌症中功能失活。因此，了解肿瘤细胞如何对 p53 激活变得不敏感至关重要。使用基于 RNAi 的遗传筛选，我们鉴定了三个调节 p53 功能的新基因。我们筛选了针对 8,000 个人类基因的 NKI shRNA 文库，以鉴定 p53 功能的调节剂。使用 shRNA 条形码技术，我们能够从复杂的混合物中快速识别活性 shRNA 载体。筛选结果的验证表明 shRNA 条形码技术可以从复杂的库中可靠地识别活性 shRNA 载体。使用这种方法，我们鉴定了三个基因：ARNTL、RBCK1 和 TNIP1，以前未知它们调节 p53 功能。重要的是，ARNTL (BMAL1) 是昼夜节律调节网络的既定组成部分。后一个发现补充了最近将昼夜节律与细胞周期和癌症联系起来的观察结果。我们发现，抑制 ARNTL 的细胞无法在 p53 激活时停滞，这与无法激活 p53 靶基因 p21CIP1 相关。我们通过功能性遗传筛选确定了 p53 通路的三个新调节因子。昼夜节律核心成分 ARNTL 的识别加强了昼夜节律与癌症之间的联系。
The p53 tumor suppressor gene is mutated in about half of human cancers, but the p53 pathway is thought to be functionally inactivated in the vast majority of cancer. Understanding how tumor cells can become insensitive to p53 activation is therefore of major importance. Using an RNAi-based genetic screen, we have identified three novel genes that regulate p53 function. We have screened the NKI shRNA library targeting 8,000 human genes to identify modulators of p53 function. Using the shRNA barcode technique we were able to quickly identify active shRNA vectors from a complex mixture. Validation of the screening results indicates that the shRNA barcode technique can reliable identify active shRNA vectors from a complex pool. Using this approach we have identified three genes, ARNTL, RBCK1 and TNIP1, previously unknown to regulate p53 function. Importantly, ARNTL (BMAL1) is an established component of the circadian regulatory network. The latter finding adds to recent observations that link circadian rhythm to the cell cycle and cancer. We show that cells having suppressed ARNTL are unable to arrest upon p53 activation associated with an inability to activate the p53 target gene p21CIP1. We identified three new regulators of the p53 pathway through a functional genetic screen. The identification of the circadian core component ARNTL strengthens the link between circadian rhythm and cancer.
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影响因子：
16
作者：
Gery, S;Komatsu, N;Koeffler, HP
通讯作者：
Koeffler, HP
影响因子：
3.5
作者：
Bayle, Julie;Lopez, Sophie;De Sepulveda, Paulo
通讯作者：
De Sepulveda, Paulo
影响因子：
64.5
作者：
ELDEIRY, WS;TOKINO, T;VOGELSTEIN, B
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VOGELSTEIN, B
影响因子：
56.9
作者：
Bunz, F;Dutriaux, A;Vogelstein, B
通讯作者：
Vogelstein, B
影响因子：
10.5
作者：
Kondratov, Roman V.;Kondratova, Anna A.;Antoch, Marina P.
通讯作者：
Antoch, Marina P.