Structural and functional analysis of the archaeal endonuclease Nob1

  • Eukaryotic ribosome biogenesis requires the concerted action of numerous ribosome assembly factors, for most of which structural and functional information is currently lacking. Nob1, which can be identified in eukaryotes and archaea, is required for the final maturation of the small subunit ribosomal RNA in yeast by catalyzing cleavage at site D after export of the preribosomal subunit into the cytoplasm. Here, we show that this also holds true for Nob1 from the archaeon Pyrococcus horikoshii, which efficiently cleaves RNA-substrates containing the D-site of the preribosomal RNA in a manganese-dependent manner. The structure of PhNob1 solved by nuclear magnetic resonance spectroscopy revealed a PIN domain common with many nucleases and a zinc ribbon domain, which are structurally connected by a flexible linker. We show that amino acid residues required for substrate binding reside in the PIN domain whereas the zinc ribbon domain alone is sufficient to bind helix 40 of the small subunit rRNA. This suggests that the zinc ribbon domain acts as an anchor point for the protein on the nascent subunit positioning it in the proximity of the cleavage site.

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Author:Thomas Veith, Roman Martin, Jan Philip WurmGND, Benjamin L. Weis, Elke Duchardt-FernerORCiD, Charlotta Safferthal, Raoul Hennig, Oliver MirusGND, Markus Bohnsack, Jens WöhnertORCiDGND, Enrico SchleiffORCiDGND
URN:urn:nbn:de:hebis:30:3-232305
URL:http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3326319/
DOI:https://doi.org/10.1093/nar/gkr1186
ISSN:1362-4962
ISSN:0305-1048
Pubmed Id:https://pubmed.ncbi.nlm.nih.gov/22156373
Parent Title (English):Nucleic acids research
Publisher:Oxford Univ. Press
Place of publication:Oxford
Document Type:Article
Language:English
Date of Publication (online):2011/12/10
Date of first Publication:2011/12/10
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2012/06/11
Volume:40
Issue:7
Page Number:16
First Page:3259
Last Page:3274
Note:
(c) The Author(s) 2011. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
HeBIS-PPN:307157253
Institutes:Biowissenschaften / Biowissenschaften
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Sammlungen:Universitätspublikationen
Sammlung Biologie / Sondersammelgebiets-Volltexte
Licence (German):License LogoCreative Commons - Namensnennung 3.0