Excision of an intact intron as a novel lariat structure during pre-mRNA splicing in vitro - PubMed (original) (raw)

Excision of an intact intron as a novel lariat structure during pre-mRNA splicing in vitro

B Ruskin et al. Cell. 1984 Aug.

Abstract

To study the mechanisms of RNA splicing we have analyzed the products generated by in vitro processing of a truncated 32P-labeled human beta-globin RNA precursor that contains the first two exons and the first intervening sequence (IVS1). Six major RNA products were detected and characterized. The first detectable RNA processing event is cleavage at the 5' GT of IVS1. Subsequently, accurately spliced RNA and the excised, intact IVS1 are simultaneously observed. The IVS1-containing RNA processing products have several unusual properties, which include: anomalous electrophoretic mobilities on polyacrylamide gels; a block to reverse transcription near the 3' end of IVS1; the presence of a nuclease-resistant component within IVS1. The block to reverse transcription and the nuclease-resistant component map to the same site near the 3' end of IVS1. The nuclease-resistant component appears to be a modified adenosine residue that contains an RNA branch. Based upon these and other structural studies we propose that the 5' end of IVS1 is joined by a 2'-5' phosphodiester linkage to the A residue in the RNAase T1 oligonucleotide ACTCTCTCTG located 28-37 nucleotides upstream from the IVS1 3' end. The IVS1 is therefore in the form of a lariat. These results imply that sequences within IVS1 actively participate in splicing.

PubMed Disclaimer

Cited by

Intron turnover is essential to the development and pathogenicity of the plant pathogenic fungus Fusarium graminearum.
Choi Y, Lee HH, Park J, Kim S, Choi S, Moon H, Shin J, Kim JE, Choi GJ, Seo YS, Son H. Choi Y, et al. Commun Biol. 2022 Oct 26;5(1):1129. doi: 10.1038/s42003-022-04111-3. Commun Biol. 2022. PMID: 36289323 Free PMC article.
The structure of the mouse parvalbumin gene.
Schleef M, Zühlke C, Jockusch H, Schöffl F. Schleef M, et al. Mamm Genome. 1992;3(4):217-25. doi: 10.1007/BF00355722. Mamm Genome. 1992. PMID: 1611216
A catalogue of splice junction and putative branch point sequences from plant introns.
Brown JW. Brown JW. Nucleic Acids Res. 1986 Dec 22;14(24):9549-59. doi: 10.1093/nar/14.24.9549. Nucleic Acids Res. 1986. PMID: 3808952 Free PMC article.
Transverse gradient gel electrophoresis: a gel system for resolving complex mixtures of lariat and linear RNA molecules.
Copertino DW, Favreau MR, Hallick RB. Copertino DW, et al. Nucleic Acids Res. 1990 Dec 25;18(24):7451-2. doi: 10.1093/nar/18.24.7451. Nucleic Acids Res. 1990. PMID: 1701880 Free PMC article.
Branch point selection in alternative splicing of tropomyosin pre-mRNAs.
Helfman DM, Ricci WM. Helfman DM, et al. Nucleic Acids Res. 1989 Jul 25;17(14):5633-50. doi: 10.1093/nar/17.14.5633. Nucleic Acids Res. 1989. PMID: 2762151 Free PMC article.

Excision of an intact intron as a novel lariat structure during pre-mRNA splicing in vitro - PubMed (original) (raw)