ArticleNucleic acids research1987
A combination of RNase H and S1 nuclease circumvents an artefact inherent to conventional S1 analysis of RNA splicing.
Article in Nucleic acids research, 1987. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed.
- During in vivo maturation of eukaryotic nuclear mRNA, splicing yields excised exon circles.Nucleic acids research · 1996Article
- Alternative splicing of a human alpha-tropomyosin muscle-specific exon: identification of determining sequences.Molecular and cellular biology · 1992Article
- Myosin light-chain 1/3 gene alternative splicing: cis regulation is based upon a hierarchical compatibility between splice sites.Molecular and cellular biology · 1990Article
- Alternative RNA splicing that is spatially regulated: generation of transcripts from the Antennapedia gene of Drosophila melanogaster with different protein-coding regions.Molecular and cellular biology · 1988Article
- Cis requirements for alternative splicing of the cardiac troponin T pre-mRNA.Nucleic acids research · 1988Article
- Use of RNase H and primer extension to analyze RNA splicing.Nucleic acids research · 1988Article
- Mechanisms of synthesis of virion proteins from the functionally bigenic late mRNAs of simian virus 40.Journal of virology · 1988Article
- Specificity of RNA maturation pathways: RNAs transcribed by RNA polymerase III are not substrates for splicing or polyadenylation.Molecular and cellular biology · 1987Article
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Authors and funding
3 authors.
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Abstract
S1 nuclease mapping is commonly used to analyze transcription and processing of unlabelled RNAs. However, the S1 protocol that appears best suited to demonstrate splicing of a particular RNA (using an intronless probe that is 5' end-labelled in the downstream exon) is not diagnostic as expected. Rather, both intron-containing RNA and intronless RNA confer protection of probe across the splice juncture. To unambiguously demonstrate correctly spliced RNAs that begin at a specific initiation site, we present a procedure in which unspliced RNA molecules are first cleaved by RNase H following annealing to an intronic DNA fragment and the remaining RNA is then subjected to S1 analysis using an intronless probe present in vast excess. Only spliced, correctly initiated transcripts can protect the probe across the splice junction and up to residue +1. This RNase H/S1 method provides a broadly applicable technique with which to demonstrate splicing and initiation of a variety of transcripts, especially ones from transfected genes that can arise both from the normal and from activated cryptic initiation sites.
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