2.50
Hdl Handle:
http://hdl.handle.net/10033/621296
Title:
Archaea box C/D enzymes methylate two distinct substrate rRNA sequences with different efficiency.
Authors:
Graziadei, Andrea; Masiewicz, Pawel; Lapinaite, Audrone; Carlomagno, Teresa ( 0000-0002-2437-2760 )
Abstract:
RNA modifications confer complexity to the 4-nucleotide polymer; nevertheless, their exact function is mostly unknown. rRNA 2'-O-ribose methylation concentrates to ribosome functional sites and is important for ribosome biogenesis. The methyl group is transferred to rRNA by the box C/D RNPs: The rRNA sequence to be methylated is recognized by a complementary sequence on the guide RNA, which is part of the enzyme. In contrast to their eukaryotic homologs, archaeal box C/D enzymes can be assembled in vitro and are used to study the mechanism of 2'-O-ribose methylation. In Archaea, each guide RNA directs methylation to two distinct rRNA sequences, posing the question whether this dual architecture of the enzyme has a regulatory role. Here we use methylation assays and low-resolution structural analysis with small-angle X-ray scattering to study the methylation reaction guided by the sR26 guide RNA fromPyrococcus furiosus We find that the methylation efficacy at sites D and D' differ substantially, with substrate D' turning over more efficiently than substrate D. This observation correlates well with structural data: The scattering profile of the box C/D RNP half-loaded with substrate D' is similar to that of the holo complex, which has the highest activity. Unexpectedly, the guide RNA secondary structure is not responsible for the functional difference at the D and D' sites. Instead, this difference is recapitulated by the nature of the first base pair of the guide-substrate duplex. We suggest that substrate turnover may occur through a zip mechanism that initiates at the 5'-end of the product.
Affiliation:
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Citation:
Archaea box C/D enzymes methylate two distinct substrate rRNA sequences with different efficiency. 2016, 22 (5):764-72 RNA
Journal:
RNA (New York, N.Y.)
Issue Date:
May-2016
URI:
http://hdl.handle.net/10033/621296
DOI:
10.1261/rna.054320.115
PubMed ID:
26925607
Type:
Article
Language:
en
ISSN:
1469-9001
Appears in Collections:
publications of te research group NMR-based structural chemistry (NBSC)

Full metadata record

DC FieldValue Language
dc.contributor.authorGraziadei, Andreaen
dc.contributor.authorMasiewicz, Pawelen
dc.contributor.authorLapinaite, Audroneen
dc.contributor.authorCarlomagno, Teresaen
dc.date.accessioned2018-02-26T09:36:56Z-
dc.date.available2018-02-26T09:36:56Z-
dc.date.issued2016-05-
dc.identifier.citationArchaea box C/D enzymes methylate two distinct substrate rRNA sequences with different efficiency. 2016, 22 (5):764-72 RNAen
dc.identifier.issn1469-9001-
dc.identifier.pmid26925607-
dc.identifier.doi10.1261/rna.054320.115-
dc.identifier.urihttp://hdl.handle.net/10033/621296-
dc.description.abstractRNA modifications confer complexity to the 4-nucleotide polymer; nevertheless, their exact function is mostly unknown. rRNA 2'-O-ribose methylation concentrates to ribosome functional sites and is important for ribosome biogenesis. The methyl group is transferred to rRNA by the box C/D RNPs: The rRNA sequence to be methylated is recognized by a complementary sequence on the guide RNA, which is part of the enzyme. In contrast to their eukaryotic homologs, archaeal box C/D enzymes can be assembled in vitro and are used to study the mechanism of 2'-O-ribose methylation. In Archaea, each guide RNA directs methylation to two distinct rRNA sequences, posing the question whether this dual architecture of the enzyme has a regulatory role. Here we use methylation assays and low-resolution structural analysis with small-angle X-ray scattering to study the methylation reaction guided by the sR26 guide RNA fromPyrococcus furiosus We find that the methylation efficacy at sites D and D' differ substantially, with substrate D' turning over more efficiently than substrate D. This observation correlates well with structural data: The scattering profile of the box C/D RNP half-loaded with substrate D' is similar to that of the holo complex, which has the highest activity. Unexpectedly, the guide RNA secondary structure is not responsible for the functional difference at the D and D' sites. Instead, this difference is recapitulated by the nature of the first base pair of the guide-substrate duplex. We suggest that substrate turnover may occur through a zip mechanism that initiates at the 5'-end of the product.en
dc.language.isoenen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.subject.meshArchaeaen
dc.subject.meshEnzymesen
dc.subject.meshMethylationen
dc.subject.meshMutationen
dc.subject.meshNucleic Acid Conformationen
dc.subject.meshRNA, Ribosomalen
dc.titleArchaea box C/D enzymes methylate two distinct substrate rRNA sequences with different efficiency.en
dc.typeArticleen
dc.contributor.departmentHelmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.en
dc.identifier.journalRNA (New York, N.Y.)en

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