Eukaryotic Box C/D methylation machinery has two non-symmetric protein assembly sites

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Authors

  • Simone Höfler
  • Peer Lukat
  • Wulf Blankenfeldt
  • Teresa Carlomagno

External Research Organisations

  • Helmholtz Centre for Infection Research (HZI)
  • Technische Universität Braunschweig
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Details

Original languageEnglish
Article number17561
JournalScientific reports
Volume11
Early online date2 Sept 2021
Publication statusPublished - Dec 2021

Abstract

Box C/D ribonucleoprotein complexes are RNA-guided methyltransferases that methylate the ribose 2’-OH of RNA. The central ‘guide RNA’ has box C and D motifs at its ends, which are crucial for activity. Archaeal guide RNAs have a second box C’/D’ motif pair that is also essential for function. This second motif is poorly conserved in eukaryotes and its function is uncertain. Conflicting literature data report that eukaryotic box C’/D’ motifs do or do not bind proteins specialized to recognize box C/D-motifs and are or are not important for function. Despite this uncertainty, the architecture of eukaryotic 2’-O-methylation enzymes is thought to be similar to that of their archaeal counterpart. Here, we use biochemistry, X-ray crystallography and mutant analysis to demonstrate the absence of functional box C’/D’ motifs in more than 80% of yeast guide RNAs. We conclude that eukaryotic Box C/D RNPs have two non-symmetric protein assembly sites and that their three-dimensional architecture differs from that of archaeal 2’-O-methylation enzymes.

ASJC Scopus subject areas

Cite this

Eukaryotic Box C/D methylation machinery has two non-symmetric protein assembly sites. / Höfler, Simone; Lukat, Peer; Blankenfeldt, Wulf et al.
In: Scientific reports, Vol. 11, 17561, 12.2021.

Research output: Contribution to journalArticleResearchpeer review

Höfler S, Lukat P, Blankenfeldt W, Carlomagno T. Eukaryotic Box C/D methylation machinery has two non-symmetric protein assembly sites. Scientific reports. 2021 Dec;11:17561. Epub 2021 Sept 2. doi: 10.1038/s41598-021-97030-y, 10.15488/11764
Höfler, Simone ; Lukat, Peer ; Blankenfeldt, Wulf et al. / Eukaryotic Box C/D methylation machinery has two non-symmetric protein assembly sites. In: Scientific reports. 2021 ; Vol. 11.
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abstract = "Box C/D ribonucleoprotein complexes are RNA-guided methyltransferases that methylate the ribose 2{\textquoteright}-OH of RNA. The central {\textquoteleft}guide RNA{\textquoteright} has box C and D motifs at its ends, which are crucial for activity. Archaeal guide RNAs have a second box C{\textquoteright}/D{\textquoteright} motif pair that is also essential for function. This second motif is poorly conserved in eukaryotes and its function is uncertain. Conflicting literature data report that eukaryotic box C{\textquoteright}/D{\textquoteright} motifs do or do not bind proteins specialized to recognize box C/D-motifs and are or are not important for function. Despite this uncertainty, the architecture of eukaryotic 2{\textquoteright}-O-methylation enzymes is thought to be similar to that of their archaeal counterpart. Here, we use biochemistry, X-ray crystallography and mutant analysis to demonstrate the absence of functional box C{\textquoteright}/D{\textquoteright} motifs in more than 80% of yeast guide RNAs. We conclude that eukaryotic Box C/D RNPs have two non-symmetric protein assembly sites and that their three-dimensional architecture differs from that of archaeal 2{\textquoteright}-O-methylation enzymes.",
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AU - Höfler, Simone

AU - Lukat, Peer

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AU - Carlomagno, Teresa

N1 - Funding Information: Open Access funding enabled and organized by Projekt DEAL. This work was funded by the Deutsche Forschun-gsgemeinschaft through a project grant to T.C. in the frame of the priority program “Chemical Biology of Natural Nucleic Acids Modifications” (CA 294/11–1).

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