Structures of RNA phosphotransferase Tpt1 reveal distinct binding modes for an RNA 2'-PO4 splice junction versus a 5'-PO4 mononucleotide.

IF 5 3区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
RNA Pub Date : 2025-06-16 DOI:10.1261/rna.080444.125
Agata Jacewicz, Masad J Damha, Stewart Shuman
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引用次数: 0

Abstract

Tpt1 is a widely distributed enzyme that removes an internal RNA 2'-phosphate by transfer to NAD+, via a two-step reaction in which: (i) the RNA 2'-PO4 attacks NAD+ to form an RNA-2'-phospho-(ADP-ribose) intermediate and expel nicotinamide; and (ii) the ADP-ribose O2″ attacks the RNA 2'-phosphodiester to form 2'-OH RNA and ADP-ribose-1″,2″-cyclic phosphate products. Tpt1 can also execute a single-step ADP-ribosyltransferase reaction at a 5'-monophosphate nucleic acid terminus that installs a 5'-phospho-ADP-ribose cap structure. Here we present crystal structures of Tpt1 bound to an RNA containing an internal 2'-PO4 mark (the substrate for the canonical Tpt1 pathway) and in a complex with 5'-AMP. We find that Tpt1 has distinct binding modes, whereby the RNA 2'-PO4 and the AMP 5'-PO4 are engaged by the same set of active site amino acids, but the 2'-PO4 nucleoside and the 5'-nucleoside occupy different sites on the enzyme.

RNA磷酸转移酶Tpt1的结构揭示了RNA 2'-PO4剪接与5'-PO4单核苷酸的不同结合模式。
Tpt1是一种广泛分布的酶,通过转移到NAD+来去除内部的RNA 2'-磷酸,通过两步反应:(i) RNA 2'- po4攻击NAD+形成RNA-2'-磷酸(adp -核糖)中间体并排出烟酰胺;(ii) adp -核糖O2”攻击RNA 2’-磷酸二酯,形成2’-OH RNA和adp -核糖1’,2’-环磷酸产物。Tpt1也可以在安装5'-磷酸- adp -核糖帽结构的5'-单磷酸核酸端执行单步adp -核糖基转移酶反应。在这里,我们展示了Tpt1与含有内部2'-PO4标记(典型Tpt1途径的底物)的RNA结合的晶体结构,并与5'-AMP复合物结合。我们发现Tpt1具有不同的结合模式,其中RNA 2'-PO4和AMP 5'-PO4由同一组活性位点氨基酸结合,但2'-PO4核苷和5'-核苷占据酶上的不同位点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
RNA
RNA 生物-生化与分子生物学
CiteScore
8.30
自引率
2.20%
发文量
101
审稿时长
2.6 months
期刊介绍: RNA is a monthly journal which provides rapid publication of significant original research in all areas of RNA structure and function in eukaryotic, prokaryotic, and viral systems. It covers a broad range of subjects in RNA research, including: structural analysis by biochemical or biophysical means; mRNA structure, function and biogenesis; alternative processing: cis-acting elements and trans-acting factors; ribosome structure and function; translational control; RNA catalysis; tRNA structure, function, biogenesis and identity; RNA editing; rRNA structure, function and biogenesis; RNA transport and localization; regulatory RNAs; large and small RNP structure, function and biogenesis; viral RNA metabolism; RNA stability and turnover; in vitro evolution; and RNA chemistry.
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