An in vivo genetic screen for genes involved in spliced leader trans-splicing indicates a crucial role for continuous de novo spliced leader RNP assembly.
An in vivo genetic screen for genes involved in spliced leader trans-splicing indicates a crucial role for continuous de novo spliced leader RNP assembly.
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DOI：
10.1093/nar/gkx500
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发表时间：
2017-08-21
影响因子：
14.9
通讯作者：
Müller B
中科院分区：
文献类型：
--
作者：
Philippe L;Pandarakalam GC;Fasimoye R;Harrison N;Connolly B;Pettitt J;Müller B
剪接前导序列（SL）反式剪接是许多真核生物基因表达的关键元件。这一过程可以说是最好的理解线虫，在秀丽隐杆线虫和猪蛔虫的生化和分子研究已经确定了关键步骤和因素。尽管如此，SL反式剪接的精确细节尚未阐明。在某种程度上，这是因为由于缺乏与该过程中的缺陷相关的特定表型，因此以前无法系统地鉴定所涉及的分子。我们在这里提出了一种新的基于GFP的报告分析，可以监测SL 1在活的C。优雅使用这种方法，我们已经确定了突变体的sna-1是有缺陷的SL反式剪接，并证明，减少功能的SNA-1，SNA-2和SUT-1，蛋白质与SL 1 RNA和相关的SmY RNA，损害SL反式剪接。我们进一步证明，Sm蛋白和pICln，SMN和Gemin 5，这是参与小核核糖核蛋白的组装，SL反式剪接有重要的作用。总之，这些结果提供了第一个在体内的证据，参与SL反式剪接的蛋白质，并表明在反式剪接反应过程中消耗的SL核糖核蛋白的连续更换是必要的有效的反式剪接。
Spliced leader (SL) trans-splicing is a critical element of gene expression in a number of eukaryotic groups. This process is arguably best understood in nematodes, where biochemical and molecular studies in Caenorhabditis elegans and Ascaris suum have identified key steps and factors involved. Despite this, the precise details of SL trans-splicing have yet to be elucidated. In part, this is because the systematic identification of the molecules involved has not previously been possible due to the lack of a specific phenotype associated with defects in this process. We present here a novel GFP-based reporter assay that can monitor SL1 trans-splicing in living C. elegans. Using this assay, we have identified mutants in sna-1 that are defective in SL trans-splicing, and demonstrate that reducing function of SNA-1, SNA-2 and SUT-1, proteins that associate with SL1 RNA and related SmY RNAs, impairs SL trans-splicing. We further demonstrate that the Sm proteins and pICln, SMN and Gemin5, which are involved in small nuclear ribonucleoprotein assembly, have an important role in SL trans-splicing. Taken together these results provide the first in vivo evidence for proteins involved in SL trans-splicing, and indicate that continuous replacement of SL ribonucleoproteins consumed during trans-splicing reactions is essential for effective trans-splicing.
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影响因子：
10.5
作者：
Evans, D;Perez, I;Blumenthal, T
通讯作者：
Blumenthal, T
影响因子：
5.3
作者：
Evans, D;Blumenthal, T
通讯作者：
Blumenthal, T
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4.5
作者：
Godfrey, Ashley C.;White, Anne E.;Duronio, Robert J.
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Duronio, Robert J.
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12.3
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影响因子：
7
作者：
Allen, Mary Ann;Hillier, LaDeana W.;Blumenthal, Thomas
通讯作者：
Blumenthal, Thomas