The AAA-ATPase NVL2 is a component of pre-ribosomal particles that interacts with the DExD/H-box RNA helicase DOB1.
The AAA-ATPase NVL2 is a component of pre-ribosomal particles that interacts with the DExD/H-box RNA helicase DOB1.
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DOI：
10.1016/j.bbrc.2006.06.017
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发表时间：
2006-08
影响因子：
3.1
通讯作者：
M. Nagahama;Takeshi Yamazoe;Y. Hara;K. Tani;A. Tsuji;M. Tagaya
中科院分区：
文献类型：
--
作者：
M. Nagahama;Takeshi Yamazoe;Y. Hara;K. Tani;A. Tsuji;M. Tagaya
核VCP/p97样蛋白2（NVL 2）是具有两个保守的ATP结合模块的伴侣样AAA-ATP酶家族的成员。我们以前的研究表明NVL 2通过与核糖体蛋白L5相互作用定位于核仁，并可能参与核糖体合成，这一过程涉及各种非核糖体因子，包括分子伴侣和RNA解旋酶。在这里，我们表明NVL 2与细胞核中的前核糖体颗粒相关。此外，我们使用酵母双杂交和免疫共沉淀试验，以确定NVL 2相互作用的蛋白质，可以深入了解NVL 2在核糖体生物合成中的功能。我们发现NVL 2与DOB 1相互作用，DOB 1是一种DExD/H-box RNA解旋酶，其酵母同源物在60 S亚基合成的后期发挥作用。DOB 1可以与NVL 2的第二个ATP结合模块突变体相互作用，该突变体对核糖体合成表现出显性负效应。相反，它不能与第一个ATP结合模块突变体相互作用，该突变体不显示显性负效应。当NVL 2的显性负突变体在细胞中过表达时，DOB 1似乎仍然与核前核糖体颗粒相关。野生型NVL 2或非显性负突变体过表达时未观察到这种积累。综上所述，我们的研究结果表明，NVL 2可能通过作为分子伴侣调节DOB 1与前核糖体颗粒的缔合/解离反应。
Nuclear VCP/p97-like protein 2 (NVL2) is a member of the chaperone-like AAA-ATPase family with two conserved ATP-binding modules. Our previous studies have shown that NVL2 is localized to the nucleolus by interacting with ribosomal protein L5 and may participate in ribosome synthesis, a process involving various non-ribosomal factors including chaperones and RNA helicases. Here, we show that NVL2 is associated with pre-ribosomal particles in the nucleus. Moreover, we used yeast two-hybrid and co-immunoprecipitation assays to identify an NVL2-interacting protein that could yield insights into NVL2 function in ribosome biogenesis. We found that NVL2 interacts with DOB1, a DExD/H-box RNA helicase, whose yeast homologue functions in a late stage of the 60S subunit synthesis. DOB1 can interact with a second ATP-binding module mutant of NVL2, which shows a dominant negative effect on ribosome synthesis. In contrast, it cannot interact with a first ATP-binding module mutant, which does not show the dominant negative effect. When the dominant negative mutant of NVL2 was overexpressed in cells, DOB1 appeared to remain associated with nuclear pre-ribosomal particles. Such accumulation was not observed upon overexpression of wild-type NVL2 or a nondominant-negative mutant. Taken together, our results suggest that NVL2 might regulate the association/dissociation reaction of DOB1 with pre-ribosomal particles by acting as a molecular chaperone.