Not all pseudouridine synthases are potently inhibited by RNA containing 5-fluorouridine
Not all pseudouridine synthases are potently inhibited by RNA containing 5-fluorouridine
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DOI：
10.1261/rna.5100104
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发表时间：
2004-02-01
期刊：
影响因子：
4.5
通讯作者：
Mueller, EG
中科院分区：
文献类型：
--
作者：
Spedaliere, CJ;Mueller, EG
已经假定含有5-氟尿苷的RNA强烈地或不可逆地抑制作用于RNA的假尿苷脱氢酶。因此，含有5-氟尿苷代替尿苷的RNA转录物已被添加到重构系统中，以便通过灭活负责其产生的假尿苷合酶来研究给定RNA中特定假尿苷残基的重要性。与含有5-氟尿苷的RNA对假尿苷合成酶的普遍抑制的假设明显矛盾的是，具有生理学上关键的真核同源物的大肠杆菌假尿苷合成酶TruB不被这样的RNA抑制。相反，含有5-氟尿苷的RNA作为底物由TruB处理。急诊另一方面，大肠杆菌假尿苷合酶RluA形成共价复合物，并被含有5-氟尿苷的RNA化学计量地抑制。我们提供了一个假设，这种不同的行为，并敦促谨慎解释结果的重建实验中，含有5-氟尿苷的RNA被假定为抑制假尿苷合酶，作为正常的功能可能会导致失败，而不是从不必要的假尿苷残基的情况下，靶向酶。
RNA containing 5-fluorouridine has been assumed to inhibit strongly or irreversibly the pseudouridine synthases that act on the RNA. RNA transcripts containing 5-fluorouridine in place of uridine have, therefore, been added to reconstituted systems in order to investigate the importance of particular pseudouridine residues in a given RNA by inactivating the pseudouridine synthase responsible for their generation. in sharp contradiction to the assumption of universal inhibition of pseudouridine synthases by RNA containing 5-fluorouridine, the Escherichia coli pseudouridine synthase TruB, which has physiologically critical eukaryotic homologs, is not inhibited by such RNA. Instead, the RNA containing 5-fluorouridine was handled as a substrate by TruB. The E. coli pseudouridine synthase RluA, on the other hand, forms a covalent complex and is inhibited stoichiometrically by RNA containing 5-fluorouridine. We offer a hypothesis for this disparate behavior and urge caution in interpreting results from reconstitution experiments in which RNA containing 5-fluorouridine is assumed to inhibit a pseudouridine synthase, as normal function may result from a failure to inactivate the targeted enzyme rather than from the absence of nonessential pseudouridine residues.