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Volumn 273, Issue 5279, 1996, Pages 1189-1195

RNA editing: A mechanism for gRNA-specified uridylate insertion into precursor mRNA

Author keywords

[No Author keywords available]

Indexed keywords

GUIDE RNA; MESSENGER RNA; PROTOZOAL RNA; RNA PRECURSOR; URIDINE PHOSPHATE; URIDINE TRIPHOSPHATE;

EID: 0029788229     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.273.5279.1189     Document Type: Article
Times cited : (164)

References (40)
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    • R. Benne, Ed. Harwood, Chichester, UK
    • S. D. Seiwert, Parasitol. Today 11, 362 (1995); K. Stuart, in RNA Editing: The Alteration of Protein Coding Sequences of RNA, R. Benne, Ed. (Harwood, Chichester, UK, 1993), p. 26; L. Simpson, D. A. Maslov. B. Blum, ibid., p. 53; R. Benne, Eur. J. Biochem. 221, 9 (1994); B. K. Adler and S. L. Hajduk, Curr. Opin. Genet. Dev. 4, 316 (1994).
    • (1993) RNA Editing: The Alteration of Protein Coding Sequences of RNA , pp. 26
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    • S. D. Seiwert, Parasitol. Today 11, 362 (1995); K. Stuart, in RNA Editing: The Alteration of Protein Coding Sequences of RNA, R. Benne, Ed. (Harwood, Chichester, UK, 1993), p. 26; L. Simpson, D. A. Maslov. B. Blum, ibid., p. 53; R. Benne, Eur. J. Biochem. 221, 9 (1994); B. K. Adler and S. L. Hajduk, Curr. Opin. Genet. Dev. 4, 316 (1994).
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    • note
    • 32P]pCp labeling.
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    • note
    • 32P]UTP and unlabeled, periodate-treated A6-eES1 were substituted for unlabeled UTP and for end-labeled A6-eES1, respectively, and reactions were scaled up twofold. After in vitro reaction and electrophoresis, RNAs with the mobility expected of edited product were excised, eluted, and precipitated with ethanol. One-half of the gel-purified samples was treated with RNase H (Gibco-BRL) in the presence of 1.6 μg of oligonucleotide anti-gA6anchor (5′-GATCTTATTCTATAACTCCAA-3′) as directed by the manufacturer. Complete RNase T1 digestion was performed at 37°C for 15 min and included 20 U of RNase T1, 50 mM tris-HCl (pH 7.4), and 2 mM EDTA.
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    • note
    • We thank members of the Stuart laboratory for helpful discussions and M. Ares, C. Mehlin, and D. Toczyski for comments on the manuscript. S.D.S. thanks J. A. Steitz for valuable insight throughout the course of this work. M.L.K. was supported in part by National Institutes of Health Biotechnology Fellowship grant GM08347. S.H. was supported by a research fellowship from the Deutsche Forschungsgemeinschaft, and this work was supported by NIH grant GM42188 to K.S.


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