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bioRxiv · 10.64898/2026.06.29.735188

18S rRNA 3’ end cleavage by the phosphorylated endoribonuclease NOB1 is interconnected with early small subunit biogenesis

Blottnitz, K.·Honemann-Capito, M.·Hackert, P.·Dybkov, O.·Lenz, C.·Bohnsack, M.·Lorenz, S.·Urlaub, H.·Schneider, C.·Bohnsack, K.
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Abstract

Processing of the precursor ribosomal RNAs (pre-rRNAs) is a key aspect of ribosomal subunit assembly that is closely coordinated with other maturation events. The ribonucleases that mediate pre-rRNA cleavages require regulation to ensure that their activities are exerted in a timely manner. Post-translational modifications can influence protein functions, and although many human ribosome assembly factors are reported to be post-translationally modified, most of these sites remain unconfirmed and functional insights are lacking. Here, we show that NOB1, the PIN domain endoribonuclease responsible for cleavage of the 3' end of the 18S rRNA, is phosphorylated within an evolutionarily conserved acidic tract that can be modified by casein kinase II in vitro. Association of NOB1 with pre-ribosomes is independent of these phosphorylations, and lack of NOB1 phosphorylation only mildly perturbs the efficiency of SSU maturation events upstream of 3' end cleavage of the 18S rRNA. Interestingly, our analyses of pre-rRNA levels in cells depleted of NOB1 or lacking its catalytic activity revealed not only accumulation of the 18SE precursor of the 18S rRNA, but also altered levels of pre-rRNAs containing 5' external transcribed spacer (ETS) sequences (43S, 26S and 30S). This suggests that lack of NOB1-mediated pre-rRNA cleavage impairs recycling of assembly factors required during early biogenesis steps, leading to altered kinetics of 5' ETS processing. Taken together these data provide new insights into the role of NOB1 during SSU biogenesis and the post-translational regulation of this ribonuclease.

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Blottnitz, K., Honemann-Capito, M., Hackert, P., Dybkov, O., Lenz, C., Bohnsack, M., Lorenz, S., Urlaub, H., Schneider, C., Bohnsack, K.. 2026-06-29. 18S rRNA 3’ end cleavage by the phosphorylated endoribonuclease NOB1 is interconnected with early small subunit biogenesis. https://doi.org/10.64898/2026.06.29.735188

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