Proteomics Uncovers Novel Components of an Interactive Protein Network Supporting RNA Export in Trypanosomes.
Proteomics Uncovers Novel Components of an Interactive Protein Network Supporting RNA Export in Trypanosomes.
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DOI：
10.1016/j.mcpro.2022.100208
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发表时间：
2022-03
期刊：
影响因子：
--
通讯作者：
Ávila AR
中科院分区：
文献类型：
--
作者：
Inoue AH;Domingues PF;Serpeloni M;Hiraiwa PM;Vidal NM;Butterfield ER;Del Pino RC;Ludwig A;Boehm C;Field MC;Ávila AR
在锥虫中，转录是多顺反子的，并且所有mRNA通过反式剪接加工，输出由非经典机制介导。虽然mRNA输出是基因调控和表达的核心，但在高等真核生物中参与mRNA输出的蛋白质的直向同源物在锥虫基因组中是可检测的，因此需要直接鉴定蛋白质组分。我们先前描述了克氏锥虫中保守的mRNA输出途径组分，包括TREX复合物的组分Sub2和外显子连接复合物（EJC）的核心组分eIF4AIII（先前为Hel45）的直系同源物。在这里，我们使用冷冻研磨和质谱法搜索这两种蛋白质的蛋白质相互作用物。TcSub2和TceIF 4AIII相互作用蛋白质群之间的显着重叠表明这两种蛋白质与相似的机制相关。我们确定了几个相互作用与保守的核心组成部分的EJC和多个额外的复合物，以及蛋白质的特定锥虫。动质体特异性蛋白质的额外免疫分离验证并扩展了超相互作用组，其能够支持从剪接到核输出和细胞质事件的RNA加工。我们还认为，只有蛋白质组学是强大的，足以揭示mRNA代谢的多个方面之间的高度连接，并揭示动质体特异性成分，创造一个独特的汞合金，以支持锥虫mRNA成熟。锥虫的基因表达由非经典机制介导。锥虫mRNA核输出系统包括独特的蛋白质到动质体。目前的工作突出了动质体特异性和保守成分的混合物。我们的数据支持一个高度耦合的mRNA成熟途径。这项工作是在完全遵守道德的情况下执行的，作者仅限于那些做出重大贡献的人。手稿是原创作品，并适当引用了他人的作品。我们在公共数据库中为适当的数据集提供原始数据。这项工作是按照社区可接受的准则和参数进行的。未造成已知危害，也未涉及人类受试者。用于生产抗体的动物使用根据机构伦理指南进行。
In trypanosomatids, transcription is polycistronic and all mRNAs are processed by trans-splicing, with export mediated by noncanonical mechanisms. Although mRNA export is central to gene regulation and expression, few orthologs of proteins involved in mRNA export in higher eukaryotes are detectable in trypanosome genomes, necessitating direct identification of protein components. We previously described conserved mRNA export pathway components in Trypanosoma cruzi, including orthologs of Sub2, a component of the TREX complex, and eIF4AIII (previously Hel45), a core component of the exon junction complex (EJC). Here, we searched for protein interactors of both proteins using cryomilling and mass spectrometry. Significant overlap between TcSub2 and TceIF4AIII-interacting protein cohorts suggests that both proteins associate with similar machinery. We identified several interactions with conserved core components of the EJC and multiple additional complexes, together with proteins specific to trypanosomatids. Additional immunoisolations of kinetoplastid-specific proteins both validated and extended the superinteractome, which is capable of supporting RNA processing from splicing through to nuclear export and cytoplasmic events. We also suggest that only proteomics is powerful enough to uncover the high connectivity between multiple aspects of mRNA metabolism and to uncover kinetoplastid-specific components that create a unique amalgam to support trypanosome mRNA maturation. Gene expression in trypanosomes is mediated by noncanonical mechanisms. Trypanosome mRNA nuclear export system comprises unique proteins to kinetoplastids. The present work highlights an amalgam of kinetoplastid-specific and conserved components. Our data support a highly coupled mRNA maturation pathway. This work was executed under full ethical compliance, and authorship is limited to those who have made significant contributions. The manuscript is original work, and works of others have been appropriately cited. We provide raw data for appropriate datasets at public databases. This work was performed standard compliant with community-acceptable guidelines and parameters. No known hazard was caused, nor any involvement of human subjects. Animal use for production of antibodies was performed under the institutional ethical guidelines.
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影响因子：
3
作者：
Chang JM;Di Tommaso P;Taly JF;Notredame C
通讯作者：
Notredame C
影响因子：
64.5
作者：
Akiyoshi B;Gull K
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作者：
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Aly 和 THO 是人类 TREX 复合物组装以及 TREX 成分与剪接 mRNA 关联所必需的
影响因子：
14.9
作者：
Chi B;Wang Q;Wu G;Tan M;Wang L;Shi M;Chang X;Cheng H
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影响因子：
4.4
作者：
Cox, Juergen;Neuhauser, Nadin;Mann, Matthias
通讯作者：
Mann, Matthias