CAPTURE of the Human U2 snRNA Genes Expands the Repertoire of Associated Factors.
CAPTURE of the Human U2 snRNA Genes Expands the Repertoire of Associated Factors.
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人类U2单链RNA基因的捕获扩展了相关因子的谱系。
DOI：
10.3390/biom12050704
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发表时间：
2022-05-14
期刊：
影响因子：
5.5
通讯作者：
Murphy, Shona
中科院分区：
文献类型：
--
作者：
Guiro, Joana;Fagbemi, Mathias;Tellier, Michael;Zaborowska, Justyna;Barker, Stephanie;Fournier, Marjorie;Murphy, Shona
为了鉴定参与人类snRNA基因转录和转录物3′端加工的因子，我们对人类细胞中串联重复的U2 snRNA基因进行了CRISPR原位亲和纯化调控元件（CAPTURE），这是deadCas 9介导的下拉。CAPTURE富集了许多预期与这些人snRNA基因相关的因子，包括RNA聚合酶II（pol II）、细胞周期蛋白依赖性激酶7（CDK 7）、负延伸因子（NELF）、Ty 5抑制因子（SPT 5）、介体23（MED 23）和整合子复合物的几个亚基。Ty 6抑制因子（SPT 6）;细胞周期蛋白K，细胞周期蛋白依赖性激酶12（CDK 12）和细胞周期蛋白依赖性激酶13（CDK 13）的伴侣;和SWI/SNF染色质重塑复合物相关的SWI/SNF相关的、基质相关的染色质调节因子（SMRC）也得到富集。几种多聚腺苷酸化因子，包括切割和多聚腺苷酸化特异性因子1（CPSF 1）、切割刺激因子1和2（CSTF 1和CSTF 2）通过U2基因CAPTURE富集。我们已经通过染色质免疫沉淀（ChIP）显示，CSTF 2-和Pcf 11和Ssu 72，也是多聚腺苷酸化因子-与人类U1和U2基因相关。ChIP-seq和ChIP-qPCR证实了SPT 6、细胞周期蛋白K和CDK 12与U2基因的关联。此外，SPT 6的敲低导致整合子复合物（INTS 3）的亚基3从U2基因中丢失，表明在snRNA基因表达中的功能作用。因此，CAPTURE扩大了与这些基因相关的转录和RNA加工因子的库，并有助于确定SPT 6的功能作用。
In order to identify factors involved in transcription of human snRNA genes and 3′ end processing of the transcripts, we have carried out CRISPR affinity purification in situ of regulatory elements (CAPTURE), which is deadCas9-mediated pull-down, of the tandemly repeated U2 snRNA genes in human cells. CAPTURE enriched many factors expected to be associated with these human snRNA genes including RNA polymerase II (pol II), Cyclin-Dependent Kinase 7 (CDK7), Negative Elongation Factor (NELF), Suppressor of Ty 5 (SPT5), Mediator 23 (MED23) and several subunits of the Integrator Complex. Suppressor of Ty 6 (SPT6); Cyclin K, the partner of Cyclin-Dependent Kinase 12 (CDK12) and Cyclin-Dependent Kinase 13 (CDK13); and SWI/SNF chromatin remodelling complex-associated SWI/SNF-related, Matrix-associated, Regulator of Chromatin (SMRC) factors were also enriched. Several polyadenylation factors, including Cleavage and Polyadenylation Specificity Factor 1 (CPSF1), Cleavage Stimulation Factors 1 and 2 (CSTF1,and CSTF2) were enriched by U2 gene CAPTURE. We have already shown by chromatin immunoprecipitation (ChIP) that CSTF2—and Pcf11 and Ssu72, which are also polyadenylation factors—are associated with the human U1 and U2 genes. ChIP-seq and ChIP-qPCR confirm the association of SPT6, Cyclin K, and CDK12 with the U2 genes. In addition, knockdown of SPT6 causes loss of subunit 3 of the Integrator Complex (INTS3) from the U2 genes, indicating a functional role in snRNA gene expression. CAPTURE has therefore expanded the repertoire of transcription and RNA processing factors associated with these genes and helped to identify a functional role for SPT6.
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影响因子：
14.9
作者：
Gene Ontology Consortium
通讯作者：
Gene Ontology Consortium
影响因子：
5.8
作者：
Guiro J;Murphy S
通讯作者：
Murphy S
影响因子：
5.3
作者：
Egloff, Sylvain;Al-Rawaf, Hadeel;Murphy, Shona
通讯作者：
Murphy, Shona
影响因子：
1.7
作者：
Bylund, L;Kytölä, S;Weber, G
通讯作者：
Weber, G
影响因子：
4.8
作者：
Egloff, Sylvain;Szczepaniak, Sylwia Anna;Murphy, Shona
通讯作者：
Murphy, Shona