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PMID: 15208448 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Interference probing of rRNA with snoRNPs: a novel approach for functional mapping of RNA in vivo.

RNA (New York, N.Y.) ·Vol. 10 ·No. 7 ·2004-07-00 ·Pages 1130-41

Liu B, Fournier MJ

Abstract

Synthesis of eukaryotic ribosomal RNAs (rRNAs) includes methylation of scores of nucleotides at the 2'-O-ribose position (Nm) by small nucleolar RNP complexes (snoRNPs). Sequence specificity is provided by the snoRNA component through base-pairing of a guide sequence with rRNA. Here, we report that methylation snoRNPs can be targeted to many new sites in yeast rRNA, by providing the snoRNA with a novel guide sequence, and that in some cases growth and translation activity are strongly impaired. Novel snoRNAs can be expressed individually or by a unique library strategy that yields guide sequences specific for a large target region. Interference effects were observed for sites in both the small and large subunits, including the reaction center region. Targeting guide RNAs to nucleotides flanking the sensitive sites caused little or no defect, indicating that methylation is responsible for the interference rather than a simple antisense effect or misguided chaperone function. To our knowledge, this is the only approach that has been used to mutagenize the backbone of rRNA in vivo.

MeSH Terms
Base Sequence DNA Probes Exons/genetics Introns/genetics Models, Molecular Molecular Sequence Data Nucleic Acid Conformation Plasmids/genetics RNA, Ribosomal/genetics RNA, Small Nucleolar/genetics Restriction Mapping
Chemicals
DNA Probes RNA, Ribosomal RNA, Small Nucleolar
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Liu Ben
Department of Biochemistry and Molecular Biology, Lederle Graduate Research Center, University of Massachusetts, Amherst, MA 01003, USA.
Fournier Maurille J
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Article Info
Journal
RNA (New York, N.Y.)
Abbr.
RNA
ISSN
1355-8382
DOI
10.1261/rna.7190104
Published
2004-07-00
Pages
1130-41
Language
English
Region
United States
NLM ID
9509184
PMCID
PMC1370603
Subset
IM
Grants
NIGMS NIH HHS · R01 GM019351 · United States
NIGMS NIH HHS · GM19351 · United States
External Links
PubMed source DOI: 10.1261/rna.7190104 PMC Full Text View journal details
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