Visualizing maturation factor extraction from the nascent ribosome by the AAA-ATPase Drg1

Prattes, Michael; Grishkovskaya, Irina; Hodirnau, Victor-Valentin; Hetzmannseder, Christina; Zisser, Gertrude; Sailer, Carolin; Kargas, Vasileios; Loibl, Mathias; Gerhalter, Magdalena; Kofler, Lisa; Warren, Alan J.; Stengel, Florian; Haselbach, David; Bergler, Helmut

Visualizing maturation factor extraction from the nascent ribosome by the AAA-ATPase Drg1

Prattes M, Grishkovskaya I, Hodirnau V-V, Hetzmannseder C, Zisser G, Sailer C, Kargas V, Loibl M, Gerhalter M, Kofler L, Warren AJ, Stengel F, Haselbach D, Bergler H. 2022. Visualizing maturation factor extraction from the nascent ribosome by the AAA-ATPase Drg1. Nature Structural & Molecular Biology. 29(9), 942–953.

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DOI

10.1038/s41594-022-00832-5

Journal Article | Published | English

Scopus indexed

Author

Prattes, Michael; Grishkovskaya, Irina; Hodirnau, Victor-Valentin^ISTA; Hetzmannseder, Christina; Zisser, Gertrude; Sailer, Carolin; Kargas, Vasileios; Loibl, Mathias; Gerhalter, Magdalena; Kofler, Lisa; Warren, Alan J.; Stengel, Florian
All

Department

Electron Microscopy Facility

Abstract

The AAA-ATPase Drg1 is a key factor in eukaryotic ribosome biogenesis that initiates cytoplasmic maturation of the large ribosomal subunit. Drg1 releases the shuttling maturation factor Rlp24 from pre-60S particles shortly after nuclear export, a strict requirement for downstream maturation. The molecular mechanism of release remained elusive. Here, we report a series of cryo-EM structures that captured the extraction of Rlp24 from pre-60S particles by Saccharomyces cerevisiae Drg1. These structures reveal that Arx1 and the eukaryote-specific rRNA expansion segment ES27 form a joint docking platform that positions Drg1 for efficient extraction of Rlp24 from the pre-ribosome. The tips of the Drg1 N domains thereby guide the Rlp24 C terminus into the central pore of the Drg1 hexamer, enabling extraction by a hand-over-hand translocation mechanism. Our results uncover substrate recognition and processing by Drg1 step by step and provide a comprehensive mechanistic picture of the conserved modus operandi of AAA-ATPases.

Keywords

Molecular Biology; Structural Biology

Publishing Year

2022

Date Published

2022-09-12

Journal Title

Nature Structural & Molecular Biology

Acknowledgement

We thank M. Fromont-Racine, A. Johnson, J. Woolford, S. Rospert, J. P. G. Ballesta and E. Hurt for supplying antibodies. The work was supported by Boehringer Ingelheim (to D. H.), the Austrian Science Foundation FWF (grants 32536 and 32977 to H. B.), the UK Medical Research Council (MR/T012412/1 to A. J. W.) and the German Research Foundation (Emmy Noether Programme STE 2517/1-1 and STE 2517/5-1 to F.S.). We thank Norberto Escudero-Urquijo, Pablo Castro-Hartmann and K. Dent, Cambridge Institute for Medical Research, for their help in cryo-EM during early phases of this project. This research was supported by the Scientific Service Units of IST Austria through resources provided by the Electron Microscopy Facility. We thank S. Keller, Institute of Molecular Biosciences (Biophysics), University Graz for support with the quantification of the SPR particle release assay. We thank I. Schaffner, University of Natural Resources and Life Sciences, Vienna for her help in early stages of the SPR experiments.

Acknowledged SSUs

Electron Microscopy Facility

Volume

Issue

Page

942-953

ISSN

1545-9993

eISSN

1545-9985

IST-REx-ID

12262

Cite this

Prattes M, Grishkovskaya I, Hodirnau V-V, et al. Visualizing maturation factor extraction from the nascent ribosome by the AAA-ATPase Drg1. Nature Structural & Molecular Biology. 2022;29(9):942-953. doi:10.1038/s41594-022-00832-5

Prattes, M., Grishkovskaya, I., Hodirnau, V.-V., Hetzmannseder, C., Zisser, G., Sailer, C., … Bergler, H. (2022). Visualizing maturation factor extraction from the nascent ribosome by the AAA-ATPase Drg1. Nature Structural & Molecular Biology. Springer Nature. https://doi.org/10.1038/s41594-022-00832-5

Prattes, Michael, Irina Grishkovskaya, Victor-Valentin Hodirnau, Christina Hetzmannseder, Gertrude Zisser, Carolin Sailer, Vasileios Kargas, et al. “Visualizing Maturation Factor Extraction from the Nascent Ribosome by the AAA-ATPase Drg1.” Nature Structural & Molecular Biology. Springer Nature, 2022. https://doi.org/10.1038/s41594-022-00832-5.

M. Prattes et al., “Visualizing maturation factor extraction from the nascent ribosome by the AAA-ATPase Drg1,” Nature Structural & Molecular Biology, vol. 29, no. 9. Springer Nature, pp. 942–953, 2022.

Prattes, Michael, et al. “Visualizing Maturation Factor Extraction from the Nascent Ribosome by the AAA-ATPase Drg1.” Nature Structural & Molecular Biology, vol. 29, no. 9, Springer Nature, 2022, pp. 942–53, doi:10.1038/s41594-022-00832-5.

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