The TCF C-clamp DNA binding domain expands the Wnt transcriptome via alternative target recognition.
The TCF C-clamp DNA binding domain expands the Wnt transcriptome via alternative target recognition.
复制标题
DOI：
10.1093/nar/gku1186
复制
发表时间：
2014-12-16
影响因子：
14.9
通讯作者：
Waterman ML
中科院分区：
文献类型：
--
作者：
Hoverter NP;Zeller MD;McQuade MM;Garibaldi A;Busch A;Selwan EM;Hertel KJ;Baldi P;Waterman ML
LEF/TCFs通过招募β-连环蛋白到基因来激活转录，从而指导WNT/β-连环蛋白信号的最后一步。古老的非脊椎动物TCFs含有两个DNA结合域，一个用于识别Wnt反应元件的高迁移率基团(WRE；5‘-CTTTGWWS-3’)和一个用于识别富含GC的Helper基序的C-钳结构域(5‘-RCCGCC-3’)。两种脊椎动物TCFs(Tcf-1/Tcf7和Tcf-4/TCF7L2)使用C-钳作为选择性剪接结构域来调节细胞周期进程，但C-钳如何影响TCF的结合和全基因组的活性尚不清楚。在这里，我们使用了带有CHIP-SEQ的多西环素诱导系统来评估C-钳如何影响全基因组范围内的人类TCF1结合。用4‘-硫代尿苷对新生RNA进行代谢脉冲标记，与RNA-SEQ结合以连接到Wnt转录组。我们发现，C-钳能够靶向更多的基因位点，以获得更强的占位和转录调节。C-夹子同时使用辅助位点和WRES进行基因靶向，但它也将TCF1靶向那些没有容易识别的正则Wres的位点。结合芯片-seq/4‘硫代尿苷-seq分析确定了新的Wnt靶基因，包括额外的细胞增殖调节因子。因此，含有TCFs异构体的C-钳是一种有效的转录调节因子，其转录组受C-钳-辅助位点相互作用的控制。
LEF/TCFs direct the final step in Wnt/β-catenin signalling by recruiting β-catenin to genes for activation of transcription. Ancient, non-vertebrate TCFs contain two DNA binding domains, a High Mobility Group box for recognition of the Wnt Response Element (WRE; 5′-CTTTGWWS-3′) and the C-clamp domain for recognition of the GC-rich Helper motif (5′-RCCGCC-3′). Two vertebrate TCFs (TCF-1/TCF7 and TCF-4/TCF7L2) use the C-clamp as an alternatively spliced domain to regulate cell-cycle progression, but how the C-clamp influences TCF binding and activity genome-wide is not known. Here, we used a doxycycline inducible system with ChIP-seq to assess how the C-clamp influences human TCF1 binding genome-wide. Metabolic pulse-labeling of nascent RNA with 4′Thiouridine was used with RNA-seq to connect binding to the Wnt transcriptome. We find that the C-clamp enables targeting to a greater number of gene loci for stronger occupancy and transcription regulation. The C-clamp uses Helper sites concurrently with WREs for gene targeting, but it also targets TCF1 to sites that do not have readily identifiable canonical WREs. The coupled ChIP-seq/4′Thiouridine-seq analysis identified new Wnt target genes, including additional regulators of cell proliferation. Thus, C-clamp containing isoforms of TCFs are potent transcriptional regulators with an expanded transcriptome directed by C-clamp-Helper site interactions.
登录
查看更多内容
影响因子：
3.7
作者：
Mokry M;Hatzis P;de Bruijn E;Koster J;Versteeg R;Schuijers J;van de Wetering M;Guryev V;Clevers H;Cuppen E
通讯作者：
Cuppen E
影响因子：
5.3
作者：
Hoverter, Nate P.;Ting, Ju-Hui;Waterman, Marian L.
通讯作者：
Waterman, Marian L.
影响因子：
64.8
作者：
Ishitani, T;Ninomiya-Tsuji, J;Matsumoto, K
通讯作者：
Matsumoto, K
影响因子：
64.8
作者：
Bhanot, P;Brink, M;Nusse, R
通讯作者：
Nusse, R
影响因子：
2.7
作者：
Klingel, Susanne;Morath, Iris;Gradl, Dietmar
通讯作者：
Gradl, Dietmar