Dominant Rio1 kinase/ATPase catalytic mutant induces trapping of late pre-40S biogenesis factors in 80S-like ribosomes.
Dominant Rio1 kinase/ATPase catalytic mutant induces trapping of late pre-40S biogenesis factors in 80S-like ribosomes.
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DOI：
10.1093/nar/gku542
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发表时间：
2014-07
影响因子：
14.9
通讯作者：
Hurt E
中科院分区：
文献类型：
--
作者：
Ferreira-Cerca S;Kiburu I;Thomson E;LaRonde N;Hurt E
在真核生物核糖体生物合成中，保守的非典型丝氨酸/苏氨酸蛋白激酶家族成员RIO激酶（Rio 1、Rio 2和Rio 3）在核糖体小亚基生物合成中起作用。Rio 2的结构分析表明，作为一个构象敏感的ATP酶，而不是一个激酶，以调节其与前40 S亚基的动态关联的作用。然而，其他RIO激酶成员在哪一步以及通过哪种机制起作用仍然难以捉摸。在这里，我们已经确定了人类Rio 1-ATP-Mg 2+复合物的晶体结构，该复合物在指示ATP酶活性的活性位点中携带磷酸天冬氨酸。酵母中基于结构的突变表明Rio 1的催化活性调节其前40 S缔合。此外，我们提供的证据表明，Rio 1协会与一个非常晚的前40 S通过其保守的C-末端结构域。此外，rio 1显性负突变体ATP水解缺陷诱导捕获后期生物合成因子的前核糖体颗粒，这原来不是前40 S，但80 S样核糖体。因此，RIO激酶折叠产生多功能ATP酶，其在Rio 1的情况下在Rio 2步骤之后被激活以调节最终40 S成熟事件之一，此时60 S亚基被募集用于最终质量控制检查。
During eukaryotic ribosome biogenesis, members of the conserved atypical serine/threonine protein kinase family, the RIO kinases (Rio1, Rio2 and Rio3) function in small ribosomal subunit biogenesis. Structural analysis of Rio2 indicated a role as a conformation-sensing ATPase rather than a kinase to regulate its dynamic association with the pre-40S subunit. However, it remained elusive at which step and by which mechanism the other RIO kinase members act. Here, we have determined the crystal structure of the human Rio1–ATP–Mg2+ complex carrying a phosphoaspartate in the active site indicative of ATPase activity. Structure-based mutations in yeast showed that Rio1's catalytic activity regulates its pre-40S association. Furthermore, we provide evidence that Rio1 associates with a very late pre-40S via its conserved C-terminal domain. Moreover, a rio1 dominant-negative mutant defective in ATP hydrolysis induced trapping of late biogenesis factors in pre-ribosomal particles, which turned out not to be pre-40S but 80S-like ribosomes. Thus, the RIO kinase fold generates a versatile ATPase enzyme, which in the case of Rio1 is activated following the Rio2 step to regulate one of the final 40S maturation events, at which time the 60S subunit is recruited for final quality control check.
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DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
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作者：
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作者：
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影响因子：
16
作者：
Ferreira-Cerca, S;Pöll, G;Milkereit, P
通讯作者：
Milkereit, P
影响因子：
16
作者：
Deloche, O;de la Cruz, J;Linder, P
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Linder, P