Proteomic analysis of rRNA-processing complex regulated by nucleolar chaperon NVL2

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AAA family proteins regulate the dissociation of molecular complexes by their ATPase activities. Nucleolar AAA-ATPase NVL2 regulates the biogenesis of 60S ribosome via the interaction with RNA helicase DOB1/MTR4. Therefore, it is suggested that there are dissociating factors regulated by NVL2 in DOB1-containing complex. However, the dissociating proteins are not yet to be identified. In this study, the experimental system that DOB1-interacting proteins could be comprehensively identified was first established by proteomics using fluorescent two-dimensional difference gel electrophoresis and mass spectrometric analysis. Using this system, SPF30 and WDR74 were identified as the dissociating-factors regulated by NVL2 from DOB1-containing complex.