A forward genetic screen in C. elegans identifies conserved residues of spliceosomal proteins PRP8 and SNRNP200/BRR2 with a role in maintaining 5' splice site identity.
A forward genetic screen in C. elegans identifies conserved residues of spliceosomal proteins PRP8 and SNRNP200/BRR2 with a role in maintaining 5' splice site identity.
复制标题
通过对 elegans 进行前向遗传筛选，发现剪接体蛋白 PRP8 和 SNRNP200/BRR2 的保守残基在维持 5'剪接位点一致性方面发挥作用。
DOI：
10.1093/nar/gkac991
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发表时间：
2022-11-11
影响因子：
14.9
通讯作者：
Zahler, Alan M.
中科院分区：
文献类型：
--
作者：
Cartwright-Acar, Catiana H.;Osterhoudt, Kenneth;Suzuki, Jessie M. N. G. L.;Gomez, Destiny R.;Katzman, Sol;Zahler, Alan M.
剪接体在组装到前体信使RNA上时经历广泛的重排。在最早的组装步骤中，U1 snRNA识别5′剪接位点。然而，U1 snRNA在组装过程中相对较早地离开剪接体，随后通过与U6 snRNA、蛋白因子PRP 8和其他组分在构建催化位点的重排过程中的相互作用来维持5′剪接位点的同一性。利用秀丽隐杆线虫的正向遗传筛选，我们已经确定了由5′ss突变引起的运动缺陷的抑制因子。在这里，我们报告了三个新的抑制等位基因从这个屏幕上，两个在PRP 8和SNRNP 200/BRR 2。这些抑制菌株的mRNASeq研究表明，它们也影响特定的天然替代5′ss，特别是抑制PRP 8 D1549 N。在SNRNP 200的非结构化N-末端的强抑制因子N18 K表明了该区域的新作用。通过检查天然基因剪接的不同变化，检查抑制子之间的双突变体，将这些新的抑制子与先前鉴定的来自酵母的剪接抑制子进行比较，并将保守的抑制子残基映射到组装人剪接体的cryoEM结构模型上，我们的结论是，在剪接体组装的多个阶段存在多种相互作用，负责维持U1 snRNA识别的初始5′ss进入催化剂核心
The spliceosome undergoes extensive rearrangements as it assembles onto precursor messenger RNAs. In the earliest assembly step, U1snRNA identifies the 5′ splice site. However, U1snRNA leaves the spliceosome relatively early in assembly, and 5′ splice site identity is subsequently maintained through interactions with U6snRNA, protein factor PRP8, and other components during the rearrangements that build the catalytic site. Using a forward genetic screen in Caenorhabditis elegans, we have identified suppressors of a locomotion defect caused by a 5′ss mutation. Here we report three new suppressor alleles from this screen, two in PRP8 and one in SNRNP200/BRR2. mRNASeq studies of these suppressor strains indicate that they also affect specific native alternative 5′ss, especially for suppressor PRP8 D1549N. A strong suppressor at the unstructured N-terminus of SNRNP200, N18K, indicates a novel role for this region. By examining distinct changes in the splicing of native genes, examining double mutants between suppressors, comparing these new suppressors to previously identified splicing suppressors from yeast, and mapping conserved suppressor residues onto cryoEM structural models of assembling human spliceosomes, we conclude that there are multiple interactions at multiple stages in spliceosome assembly responsible for maintaining the initial 5′ss identified by U1snRNA for entry into the catalytic core.
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影响因子：
10.5
作者：
Absmeier E;Wollenhaupt J;Mozaffari-Jovin S;Becke C;Lee CT;Preussner M;Heyd F;Urlaub H;Lührmann R;Santos KF;Wahl MC
通讯作者：
Wahl MC
影响因子：
1.2
作者：
Cingolani, Pablo;Platts, Adrian;Ruden, Douglas M.
通讯作者：
Ruden, Douglas M.
影响因子：
4.3
作者：
Absmeier, Eva;Becke, Christian;Wahl, Markus C.
通讯作者：
Wahl, Markus C.
DOI：
10.1073/pnas.1819020116
发表时间：
2019-02-05
影响因子：
11.1
作者：
Mayerle, Megan;Yitiz, Samira;Zahler, Alan M.
通讯作者：
Zahler, Alan M.
影响因子：
14.9
作者：
CORPET, F
通讯作者：
CORPET, F