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Bacterial Expression System> | DNA encoding for the D1/D2 Walker B variant of C. thermophilum Rix7 was codon optimized for E. coli expression (GenScript) and inserted into the pet24b expression plasmid containing a C-terminal hexa-histidine tag (Supplementary Table 2). | Get A Quote |
Rix7 is an essential type II AAA-ATPase required for the formation of the large ribosomal subunit. Rix7 has been proposed to utilize the power of ATP hydrolysis to drive the removal of assembly factors from pre-60S particles, but the mechanism of release is unknown. Rix7's mammalian homolog, NVL2 has been linked to cancer and mental illness disorders, highlighting the need to understand the molecular mechanisms of this essential machine. Here we report the cryo-EM reconstruction of the tandem AAA domains of Rix7 which form an asymmetric stacked homohexameric ring. We trapped Rix7 with a polypeptide in the central channel, revealing Rix7's role as a molecular unfoldase. The structure establishes that type II AAA... More