Mammalian nuclear TRUB1, mitochondrial TRUB2, and cytoplasmic PUS10 produce conserved pseudouridine 55 in different sets of tRNA.
Mammalian nuclear TRUB1, mitochondrial TRUB2, and cytoplasmic PUS10 produce conserved pseudouridine 55 in different sets of tRNA.
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DOI：
10.1261/rna.076810.120
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发表时间：
2021-01
期刊：
影响因子：
--
通讯作者：
Gupta R
中科院分区：
文献类型：
--
作者：
Mukhopadhyay S;Deogharia M;Gupta R
大多数哺乳动物细胞质trna分别在54位和55位含有核糖嘧啶(T)和假尿嘧啶（Ψ）。然而，一些trna在两个位置都含有Ψ。一些Ψ54-containing trna作为逆转录病毒DNA合成的引物。这些trna的Ψ54是由PUS10产生的，它也可以合成Ψ55。另外两种酶TRUB1和TRUB2也能产生Ψ55。通过对重组蛋白和野生型和特异性Ψ55合成酶敲低细胞亚细胞提取物处理的tRNA进行近邻分析，我们确定，虽然TRUB1、PUS10和TRUB2都具有tRNA Ψ55合成酶活性，但它们对tRNA的结构要求不同。此外，这些活性主要分别存在于细胞核、细胞质和线粒体中，这表明Ψ55合成酶活性存在区隔性。TRUB1产生大多数伸长体trna的Ψ55，而细胞质PUS10产生含有Ψ54Ψ55的trna的Ψs。PUS10的核同工异构体具有催化活性，特异性结合未修饰的U54U55版本的Ψ54Ψ55-containing trna，以及含有a54u55的tRNAiMet。这种结合抑制了trub1介导的U55在细胞核中的Ψ55转化。因此，Ψ54Ψ55-containing trna的U54U55被细胞质PUS10修饰。核PUS10不结合T54Ψ55-和A54Ψ55-containing伸长体trna的U55版本。因此，TRUB1能够在这些trna中产生Ψ55。综上所述，TRUB1和PUS10的tRNA Ψ55合成酶活性不是冗余的，而是区隔的，作用于不同的tRNA组。这种划分的意义需要进一步研究。
Most mammalian cytoplasmic tRNAs contain ribothymidine (T) and pseudouridine (Ψ) at positions 54 and 55, respectively. However, some tRNAs contain Ψ at both positions. Several Ψ54-containing tRNAs function as primers in retroviral DNA synthesis. The Ψ54 of these tRNAs is produced by PUS10, which can also synthesize Ψ55. Two other enzymes, TRUB1 and TRUB2, can also produce Ψ55. By nearest-neighbor analyses of tRNAs treated with recombinant proteins and subcellular extracts of wild-type and specific Ψ55 synthase knockdown cells, we determined that while TRUB1, PUS10, and TRUB2 all have tRNA Ψ55 synthase activities, they have different tRNA structural requirements. Moreover, these activities are primarily present in the nucleus, cytoplasm, and mitochondria, respectively, suggesting a compartmentalization of Ψ55 synthase activity. TRUB1 produces the Ψ55 of most elongator tRNAs, but cytoplasmic PUS10 produces both Ψs of the tRNAs with Ψ54Ψ55. The nuclear isoform of PUS10 is catalytically inactive and specifically binds the unmodified U54U55 versions of Ψ54Ψ55-containing tRNAs, as well as the A54U55-containing tRNAiMet. This binding inhibits TRUB1-mediated U55 to Ψ55 conversion in the nucleus. Consequently, the U54U55 of Ψ54Ψ55-containing tRNAs are modified by the cytoplasmic PUS10. Nuclear PUS10 does not bind the U55 versions of T54Ψ55- and A54Ψ55-containing elongator tRNAs. Therefore, TRUB1 is able to produce Ψ55 in these tRNAs. In summary, the tRNA Ψ55 synthase activities of TRUB1 and PUS10 are not redundant but rather are compartmentalized and act on different sets of tRNAs. The significance of this compartmentalization needs further study.
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影响因子：
5.3
作者：
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