A novel, essential trans-splicing protein connects the nematode SL1 snRNP to the CBC-ARS2 complex.
A novel, essential trans-splicing protein connects the nematode SL1 snRNP to the CBC-ARS2 complex.
复制标题
DOI：
10.1093/nar/gkac534
复制
发表时间：
2022-07-22
影响因子：
14.9
通讯作者：
Mueller, Berndt
中科院分区：
文献类型：
--
作者：
Fasimoye, Rotimi Yemi;Spencer, Rosie Elizabeth Barker;Soto-Martin, Eva;Eijlers, Peter;Elmassoudi, Haitem;Brivio, Sarah;Mangana, Carolina;Sabele, Viktorija;Rechtorikova, Radoslava;Wenzel, Marius;Connolly, Bernadette;Pettitt, Jonathan;Mueller, Berndt
剪接前导序列的反式剪接对于许多真核生物的基因表达是必不可少的。为了阐明这一过程的分子机制，我们对秀丽隐杆线虫主要剪接前导序列snRNP（SL 1 snRNP）相关分子进行了测序，该序列提供了剪接前导序列，取代了大多数前体mRNA的5′非翻译区。使用由CRISPR介导的基因组工程创建的SL 1 snRNP蛋白SNA-1的GFP标记版本，我们通过RIP-Seq和质谱法免疫沉淀并鉴定RNA和蛋白质组分。这揭示了SL 1 snRNP的组成，并确定了与剪接体组分PRP-8和PRP-19的关联。值得注意的是，我们确定了一种新的，线虫特异性蛋白所需的SL 1反式剪接，我们指定SNA-3。SNA-3是一种必需的核蛋白，具有三个NADAR结构域，其功能尚不清楚。NADAR结构域中关键残基的突变使蛋白失活，表明结构域功能是活性所必需的。SNA-3与CBC-ARS 2复合物和其他参与RNA代谢的因子（包括SUT-1蛋白）通过酵母双杂交试验、定位研究和免疫沉淀揭示的RNA或蛋白质介导的接触相互作用。我们的数据与SNA-3在协调反式剪接与靶前mRNA转录或反式剪接反应的Y分支产物的加工中的作用是相容的。
Spliced leader trans-splicing is essential for gene expression in many eukaryotes. To elucidate the molecular mechanism of this process, we characterise the molecules associated with the Caenorhabditis elegans major spliced leader snRNP (SL1 snRNP), which donates the spliced leader that replaces the 5′ untranslated region of most pre-mRNAs. Using a GFP-tagged version of the SL1 snRNP protein SNA-1 created by CRISPR-mediated genome engineering, we immunoprecipitate and identify RNAs and protein components by RIP-Seq and mass spectrometry. This reveals the composition of the SL1 snRNP and identifies associations with spliceosome components PRP-8 and PRP-19. Significantly, we identify a novel, nematode-specific protein required for SL1 trans-splicing, which we designate SNA-3. SNA-3 is an essential, nuclear protein with three NADAR domains whose function is unknown. Mutation of key residues in NADAR domains inactivates the protein, indicating that domain function is required for activity. SNA-3 interacts with the CBC-ARS2 complex and other factors involved in RNA metabolism, including SUT-1 protein, through RNA or protein-mediated contacts revealed by yeast two-hybrid assays, localisation studies and immunoprecipitations. Our data are compatible with a role for SNA-3 in coordinating trans-splicing with target pre-mRNA transcription or in the processing of the Y-branch product of the trans-splicing reaction.
登录
查看更多内容
影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
影响因子：
--
作者：
Huang G;de Jesus B;Koh A;Blanco S;Rettmann A;DeMott E;Sylvester M;Ren C;Meng C;Waterland S;Rhodes A;Alicea P;Flynn A;Dickinson DJ;Doonan R
通讯作者：
Doonan R
DOI：
10.1261/rna.076414.120
发表时间：
2020-12
期刊：
RNA (New York, N.Y.)
影响因子：
--
作者：
Wenzel M;Johnston C;Müller B;Pettitt J;Connolly B
通讯作者：
Connolly B
影响因子：
4.4
作者：
Cox, Juergen;Neuhauser, Nadin;Mann, Matthias
通讯作者：
Mann, Matthias
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y