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bioRxiv · 10.1101/2024.11.02.621631

Pre-rRNA spatial distribution and functional organization of the nucleolus

Pan, Y.-H.·Shan, L.·Yang, Z.-H.·Zhang, Y.-Y.·Zhang, Y.·Cao, S.-M.·Zhang, J.·Yang, L.·Chen, L.-L.
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Abstract

The self-organized nucleolus is where the pre-ribosomal RNA (pre-rRNA) processing and pre-ribosomal ribonucleoprotein (RNP) assembly take place. Here we present the spatiotemporal distribution of pre-rRNA intermediates in the human nucleolus, revealing a segregated spatial distribution of small subunit (SSU) and large subunit (LSU) precursors. Notably, the 5 external transcribed spacer (5 ETS)-containing SSU pre-rRNAs are retained across FC-DFC-PDFC regions, while the internal transcribed spacer 2 (ITS2)-containing LSU pre-rRNAs move to PDFC-GC regions for processing. Inhibiting 5 ETS processing disrupts the SSU pre-rRNA distribution and the nested FC-DFC sub-nucleolar structure. Cells in amniotes possess a multi-layered nucleolus, whereas anamniotes have only a bipartite structure with a merged FC-DFC. Kinetic labeling of pre-rRNA outflow shows a 7-fold higher pace in tetrapartite nucleoli over that in bipartite nucleoli, indicating that the emergence of the nested FC-DFC may facilitate an efficient SSU pre-rRNA processing over the course of evolution. Collectively, depicting the spatiotemporal distribution of pre-rRNAs reveals a key role of processing steps in organizing the multi-layered nucleolus and suggests a possible evolutionary advantage of the multi-layered structure in amniotes.

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Pan, Y.-H., Shan, L., Yang, Z.-H., Zhang, Y.-Y., Zhang, Y., Cao, S.-M., Zhang, J., Yang, L., Chen, L.-L.. 2024-11-03. Pre-rRNA spatial distribution and functional organization of the nucleolus. https://doi.org/10.1101/2024.11.02.621631

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