Proteomic analysis of in vivo-assembled pre-mRNA splicing complexes expands the catalog of participating factors.
Proteomic analysis of in vivo-assembled pre-mRNA splicing complexes expands the catalog of participating factors.
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DOI：
10.1093/nar/gkm347
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发表时间：
2007
影响因子：
14.9
通讯作者：
Stevens SW
中科院分区：
文献类型：
--
作者：
Chen YI;Moore RE;Ge HY;Young MK;Lee TD;Stevens SW
先前的前mRNA加工复合物的组成研究已经在体外对含有单个内含子的合成前mRNA进行。为了提供与前体mRNA剪接装置相关的多肽的更全面的列表，我们已经确定了活细胞中大量前体mRNA加工装置的组成。我们从人和鸡细胞中纯化了内源性核前mRNA加工复合物，其包含大量（>200S）的超剪接体（也称为多剪接体）。正如预期的那样，RNA组分包括前mRNA和五种剪接体snRNA的异质混合物。除了已知的前体mRNA剪接因子、5′端结合因子、3′端加工因子、mRNA输出因子、hnRNP和其他RNA结合蛋白外，通过质谱鉴定的蛋白质组分包括RNA腺苷脱氨酶和几种新的因子。有趣的是，我们纯化的超剪接体还含有许多结构蛋白，核孔蛋白，染色质重塑因子和几种新的蛋白质，这些蛋白质在体外组装的剪接复合物中不存在。这些体内分析使与前mRNA相关的因子总数远远超过300个，并且代表了迄今为止前mRNA加工机制的最全面的分析。
Previous compositional studies of pre-mRNA processing complexes have been performed in vitro on synthetic pre-mRNAs containing a single intron. To provide a more comprehensive list of polypeptides associated with the pre-mRNA splicing apparatus, we have determined the composition of the bulk pre-mRNA processing machinery in living cells. We purified endogenous nuclear pre-mRNA processing complexes from human and chicken cells comprising the massive (>200S) supraspliceosomes (a.k.a. polyspliceosomes). As expected, RNA components include a heterogeneous mixture of pre-mRNAs and the five spliceosomal snRNAs. In addition to known pre-mRNA splicing factors, 5′ end binding factors, 3′ end processing factors, mRNA export factors, hnRNPs and other RNA binding proteins, the protein components identified by mass spectrometry include RNA adenosine deaminases and several novel factors. Intriguingly, our purified supraspliceosomes also contain a number of structural proteins, nucleoporins, chromatin remodeling factors and several novel proteins that were absent from splicing complexes assembled in vitro. These in vivo analyses bring the total number of factors associated with pre-mRNA to well over 300, and represent the most comprehensive analysis of the pre-mRNA processing machinery to date.
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影响因子：
4.8
作者：
Blevins, MB;Smith, AM;Powers, MA
通讯作者：
Powers, MA
影响因子：
16
作者：
Gilbert, W;Guthrie, C
通讯作者：
Guthrie, C
影响因子：
10.5
作者：
BENNETT, M;MICHAUD, S;REED, R
通讯作者：
REED, R
DOI：
10.1016/s1044-0305(97)00282-1
发表时间：
1998-03-01
影响因子：
3.2
作者：
Davis, MT;Lee, TD
通讯作者：
Lee, TD
DOI：
10.1073/pnas.94.22.11798
发表时间：
1997-10-28
影响因子：
11.1
作者：
Arenas, JE;Abelson, JN
通讯作者：
Abelson, JN