Human NOP2/NSUN1 regulates ribosome biogenesis through non-catalytic complex formation with box C/D snoRNPs

5-Methylcytosine (m(5)C) is a base modification broadly found on various RNAs in the human transcriptome. In eukaryotes, m(5)C is catalyzed by enzymes of the NSUN family composed of seven human members (NSUN1-7). NOP2/NSUN1 has been primarily characterized in budding yeast as an essential ribosome biogenesis factor required for the deposition of m(5)C on the 25S ribosomal RNA (rRNA). Although human NOP2/NSUN1 has been known to be an oncogene overexpressed in several types of cancer, its functions and substrates remain poorly characterized. Here, we used a miCLIP-seq approach to identify human NOP2/NSUN1 RNA substrates. Our analysis revealed that NOP2/NSUN1 catalyzes the deposition of m(5)C at position 4447 on the 28S rRNA. We also find that NOP2/NSUN1 binds to the 5 ' ETS region of the pre-rRNA transcript and regulates pre-rRNA processing through non-catalytic complex formation with box C/D snoRNAs. We provide evidence that NOP2/NSUN1 facilitates the recruitment of U3 and U8 snoRNAs to pre-90S ribosomal particles and their stable assembly into snoRNP complexes. Remarkably, expression of both WT and catalytically inactive NOP2/NSUN1 in knockdown background rescues the rRNA processing defects and the stable assembly of box C/D snoRNP complexes, suggesting that NOP2/NSUN1-mediated deposition of m(5)C on rRNA is not required for ribosome synthesis.

Automated summary

NOP2/NSUN1 catalyzes the deposition of m(5)C at position 4447 on the 28S rRNA, and regulates pre-rRNA processing through non-catalytic complex formation with box C/D snoRNAs. Additionally, NOP2/NSUN1 facilitates the recruitment and stable assembly of U3 and U8 snoRNAs into snoRNP complexes.