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PMID: 10773075 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Unique structural and stabilizing roles for the individual pseudouridine residues in the 1920 region of Escherichia coli 23S rRNA.

Nucleic acids research ·Vol. 28 ·No. 10 ·2000-05-15 ·Pages 2075-83

Meroueh M, Grohar PJ, Qiu J, SantaLucia J, Scaringe SA, Chow CS

Abstract

The synthesis of a 5'-O-BzH-2'- O -ACE-protected pseudouridine phosphoramidite is reported [BzH, benzhydryloxy-bis(trimethylsilyloxy)silyl; ACE, bis(2-acetoxyethoxy)methyl]. The availability of the phosphoramidite allows for reliable and efficient syntheses of hairpin RNAs containing single or multiple pseudouridine modifications in the stem or loop regions. Five 19-nt hairpin RNAs representing the 1920-loop region (G(1906)-C(1924)) of Escherichia coli 23S rRNA were synthesized with pseudouridine residues located at positions 1911, 1915 and 1917. Thermodynamic parameters, circular dichroism spectra and NMR data are presented for all five RNAs. Overall, three different structural contexts for the pseudouridine residues were examined and compared with the unmodified RNA. Our main findings are that pseudouridine modifications exhibit a range of effects on RNA stability and structure, depending on their locations. More specifically, pseudouridines in the single-stranded loop regions of the model RNAs are slightly destabilizing, whereas a pseudo-uridine at the stem-loop junction is stabilizing. Furthermore, the observed effects on stability are approximately additive when multiple pseudouridine residues are present. The possible relationship of these results to RNA function is discussed.

MeSH Terms
Base Sequence Circular Dichroism Escherichia coli/genetics Models, Molecular Nuclear Magnetic Resonance, Biomolecular Nucleic Acid Conformation Oligoribonucleotides/chemical synthesis,chemistry Organophosphorus Compounds/chemical synthesis Pseudouridine/analogs & derivatives,chemical synthesis,chemistry RNA, Bacterial/chemistry RNA, Ribosomal, 23S/chemistry Thermodynamics
Chemicals
5'-O-(benzhydryloxybis(trimethylsilyloxy)silyl)-2'-O-(bis(2-acetoxyethoxy)methyl)pseudouridine-3'-(methyl-N,N-diisopropylphosphoramidite) Oligoribonucleotides Organophosphorus Compounds RNA, Bacterial RNA, Ribosomal, 23S Pseudouridine
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Meroueh M
Department of Chemistry, Wayne State University, Detroit, MI 48202, USA.
Grohar P J
Qiu J
SantaLucia J
Scaringe S A
Chow C S
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/28.10.2075
Published
2000-05-15
Pages
2075-83
Language
English
Region
England
NLM ID
0411011
PMCID
PMC105375
Subset
IM
Grants
NIGMS NIH HHS · R01 GM054632 · United States
NIGMS NIH HHS · R29 GM054632 · United States
NIGMS NIH HHS · GM56044 · United States
NIGMS NIH HHS · GM54632 · United States
NIGMS NIH HHS · GM55745 · United States
External Links
PubMed source DOI: 10.1093/nar/28.10.2075 PMC Full Text View journal details
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