Structural and biochemical characterization of the 3'-5' tRNA splicing ligases.

IF 4 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Journal of Biological Chemistry Pub Date : 2025-04-10 DOI:10.1016/j.jbc.2025.108506

Sebastian Chamera,Weronika Zajko,Mariusz Czarnocki-Cieciura,Marcin Jaciuk,Łukasz Koziej,Jakub Nowak,Krzysztof Wycisk,Małgorzata Sroka,Andrzej Chramiec-Głąbik,Mirosław Śmietański,Filip Gołębiowski,Marcin Warmiński,Jacek Jemielity,Sebastian Glatt,Marcin Nowotny

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引用次数: 0

Abstract

In archaea and metazoa, tRNA exons are ligated by the RNA ligases RtcB and RTCB, respectively. The metazoan RTCB forms a stable complex with four additional subunits, DDX1, FAM98B, CGI99 and ASHWIN. The role and assembly of these four components remain elusive. Furthermore, we lack structural information of how RNA substrates are recognized by 3'-5' tRNA ligases. Here, we use thiol-based chemical cross-linking to confirm the involvement of specific residues of RtcB in RNA binding and we present a cryo-electron microscopy structure of the purified five-subunit D. rerio tRNA ligase complex. The structure implies that the DDX1 helicase module is mobile and can modulate the activity of RTCB. Taken together, the presented results enhance our mechanistic understanding of RNA binding by 3'-5' tRNA splicing ligases and architecture of the metazoan tRNA ligase complex.

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3'-5' tRNA剪接连接酶的结构和生化特性。

在古生菌和元古宙中，tRNA 外显子分别由 RNA 连接酶 RtcB 和 RTCB 连接。元古宙的 RTCB 与另外四个亚基 DDX1、FAM98B、CGI99 和 ASHWIN 形成稳定的复合体。这四种成分的作用和组装仍然难以捉摸。此外，我们还缺乏有关 3'-5' tRNA 连接酶如何识别 RNA 底物的结构信息。在这里，我们利用基于硫醇的化学交联来证实 RtcB 的特定残基参与了 RNA 结合，并展示了纯化的 D. rerio tRNA 连接酶五亚基复合物的冷冻电镜结构。该结构意味着 DDX1 螺旋酶模块是可移动的，并能调节 RTCB 的活性。综上所述，这些结果加深了我们对 3'-5' tRNA 连接酶结合 RNA 的机理和元动物 tRNA 连接酶复合物结构的理解。

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来源期刊

Journal of Biological Chemistry Biochemistry, Genetics and Molecular Biology-Biochemistry

自引率

4.20%

发文量

1233

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