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PMID: 6164994 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A sequence from Drosophila melanogaster 18S rRNA bearing the conserved hypermodified nucleoside am psi: analysis by reverse transcription and high-performance liquid chromatography.

Nucleic acids research ·Vol. 9 ·No. 7 ·1981-04-10 ·Pages 1723-41

Youvan DC, Hearst JE

Abstract

The naturally occurring modified nucleoside 3-[3-amino-3-carboxypropyl]-1-methylpseudouridine (abbreviated am psi) is found in eukaryotic 18S rRNA. We localized am psi to sequence resolution in D. melanogaster 18S rRNA. This hypermodified base causes an absolute stop in cDNA elongation. The RNA sequence bearing am psi was determined by dideoxy-sequencing with reverse transcriptase. The rDNA coding for this part of the 18S rRNA was sequenced by the Maxam-Gilbert method. Together these two sequencing methods can be used to position the cDNA stop (am psi) in the rRNA sequence. Chemical evidence for the existence of am psi in this RNA sequence was obtained by high-performance liquid chromatography (HPLC) of 18S rRNA nucleosides from radioactive-labeled cells. L-[2-14C] methionine will selectively label am psi in eukaryotic 18S rRNA. Using HPLC, we found a single 14C-labeled nucleotide in digests of 18S rRNA. This nucleotide is in the RNA sequence bearing the cDNA stop since a restriction fragment which hybridizes to this sequence protects the modified base from RNase T1 digestion.

MeSH Terms

Animals Base Sequence Chromatography, High Pressure Liquid Cloning, Molecular Drosophila melanogaster/genetics Nucleic Acid Hybridization Plasmids RNA, Ribosomal/genetics RNA-Directed DNA Polymerase/metabolism Transcription, Genetic Uracil Nucleotides/analysis Uridine Monophosphate/analogs & derivatives,analysis

Chemicals

RNA, Ribosomal Uracil Nucleotides 3-(3-amino-3-carboxypropyl)-1-methylpseudouridine Uridine Monophosphate RNA-Directed DNA Polymerase

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Youvan D C

Hearst J E

References (23)

23 references, click to expand

Complete nucleotide sequence of a 16S ribosomal RNA gene from Escherichia coli.
Proc Natl Acad Sci U S A. 1978 Oct;75(10):4801-5 PMID: 368799
Partial mapping of methylated sequences in Xenopus laevis ribosomal RNA by preparative hybridization to cloned fragments of ribosomal DNA.
Nucleic Acids Res. 1979 Mar;6(3):817-30 PMID: 108668
Studies on the methylation of cytoplasmic ribosomal RNA from cultured higher plant cells.
Eur J Biochem. 1979 Jul;98(1):203-14 PMID: 223845
Reverse transcriptase pauses at N2-methylguanine during in vitro transcription of Escherichia coli 16S ribosomal RNA.
Proc Natl Acad Sci U S A. 1979 Aug;76(8):3751-4 PMID: 91169
A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
Nucleic Acids Res. 1979 Nov 24;7(6):1513-23 PMID: 388356
Comparison of nucleotide sequences of large T1 ribonuclease fragments of 18S ribosomal RNA of rat and chicken.
Nucleic Acids Res. 1979 Nov 10;7(5):1131-5 PMID: 117430
Prediction of the electrophoretic mobilities of nucleotides on neutral paper.
Anal Biochem. 1979 Sep 15;98(1):8-12 PMID: 44439
Major and modified nucleosides in tRNA hydrolysates by high-performance liquid chromatography.
J Chromatogr. 1979 May 21;173(2):281-98 PMID: 94916
Chromatography of nucleosides.
J Chromatogr. 1980 Jan 25;188(1):129-47 PMID: 7380928
Improved methods for maximizing expression of a cloned gene: a bacterium that synthesizes rabbit beta-globin.
Cell. 1980 Jun;20(2):543-53 PMID: 6248249
Sequence of the 3'-terminal portion of Drosophila melanogaster 18 S rRNA and of the adjoining spacer: comparison with corresponding prokaryotic and eukaryotic sequences.
FEBS Lett. 1980 Aug 11;117(1):227-31 PMID: 6250894
The structure of the yeast ribosomal RNA genes. I. The complete nucleotide sequence of the 18S ribosomal RNA gene from Saccharomyces cerevisiae.
Nucleic Acids Res. 1980 Dec 11;8(23):5779-94 PMID: 7008030
The isolation from ribonucleic acid of substituted uridines containing alpha-aminobutyrate moieties derived from methionine.
Biochim Biophys Acta. 1974 Apr 27;349(1):61-77 PMID: 11400439
Sequence analysis of specific areas of the 16S and 23S ribosomal RNAs.
Nature. 1968 Jul 20;219(5151):236-8 PMID: 4299552
In vitro culture of Drosophila melanogaster embryonic cells.
In Vitro. 1970 Nov-Dec;6(3):162-72 PMID: 5002052
Presence of a hypermodified nucleotide in HeLa cell 18 S and Saccharomyces carlsbergensis 17 S ribosomal RNAs.
FEBS Lett. 1975 Nov 1;59(1):60-3 PMID: 1225623
A new method for sequencing DNA.
Proc Natl Acad Sci U S A. 1977 Feb;74(2):560-4 PMID: 265521
Sequences homologous to ribosomal insertions occur in the Drosophila genome outside the nucleolus organizer.
Proc Natl Acad Sci U S A. 1977 Oct;74(10):4233-7 PMID: 412186
DNA sequencing with chain-terminating inhibitors.
Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
Biosynthesis of a hypermodified nucleotide in Saccharomyces carlsbergensis 17S and HeLa-cell 18S ribosomal ribonucleic acid.
Biochem J. 1978 Jan 1;169(1):71-7 PMID: 629754
Structure of psoralen-crosslinked ribosomal RNA from Drosophila melanogaster.
Proc Natl Acad Sci U S A. 1978 Apr;75(4):1642-6 PMID: 417342
Conservation of the primary structure at the 3' end of 18S rRNA from eucaryotic cells.
Cell. 1978 Mar;13(3):551-63 PMID: 77738
Processing steps and methylation in the formation of the ribosomal RNA of cultured Drosophila cells.
J Mol Biol. 1978 May 15;121(2):219-38 PMID: 660652

Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/9.7.1723

Published

1981-04-10

Pages

1723-41

Language

English

Region

England

NLM ID

0411011

PMCID

PMC326793

Subset

Grants

NIEHS NIH HHS · ES 07075 · United States

Databases

GENBANK

K01286, K01287

External Links

PubMed source DOI: 10.1093/nar/9.7.1723 PMC Full Text View journal details

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A sequence from Drosophila melanogaster 18S rRNA bearing the conserved hypermodified nucleoside am psi: analysis by reverse transcription and high-performance liquid chromatography.

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