Antisense RNA inhibits splicing of pre-mRNA in vitro

Munroe, S. H. (August 1988) Antisense RNA inhibits splicing of pre-mRNA in vitro. Embo J, 7 (8). pp. 2523-32. ISSN 0261-4189

URL: http://www.ncbi.nlm.nih.gov/pubmed/2461296
DOI: 10.1002/j.1460-2075.1988.tb03100.x

Abstract

Antisense RNAs complementary to human beta-globin pre-mRNA or to a chimeric globin/adenovirus E2a pre-mRNA specifically and efficiently inhibit pre-mRNA splicing in vitro. The level of inhibition depends on the length, position and concentration of the antisense RNA relative to the pre-mRNA substrate. Antisense RNAs complementary to sequences greater than 80 nucleotides downstream of the globin 3' splice site inhibit at least as efficiently as those extending across the splice sites. Thus splicing is sensitive to perturbations involving exon sequences some distance from the splice sites. Inhibition is mediated by factors which affect the annealing of antisense and substrate RNAs. Direct analysis of RNA duplex formation demonstrates the presence of an activity in HeLa cell nuclear extract which promotes the rapid annealing of complementary RNAs in an ATP-independent manner. Both annealing and inhibition are greatly reduced when antisense RNA is added to the splicing reaction greater than or equal to 5 min after substrate. This result may reflect a transition between an open structure, in which annealing of antisense RNA with pre-mRNA is facilitated, and a closed complex in which pre-mRNA is sequestered at an early stage of spliceosome assembly.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > pre-mRNA
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNA splicing
CSHL Authors:
Communities: CSHL labs
Depositing User: Gail Sherman
Date: 1 August 1988
Date Deposited: 25 Sep 2017 19:20
Last Modified: 25 Sep 2017 19:20
PMCID: PMC457123
Related URLs:
URI: http://repository.cshl.edu/id/eprint/35189

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