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Molecular and Cellular Biology, December 1998, p. 7510-7520, Vol. 18, No. 12
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

The Pre-mRNA 5' Cap Determines Whether U6 Small Nuclear RNA Succeeds U1 Small Nuclear Ribonucleoprotein Particle at 5' Splice Sites

Laura O'Mullane and Ian C. Eperon*

Department of Biochemistry, University of Leicester, Leicester LE1 7RH, United Kingdom

Received 4 June 1998/Returned for modification 10 August 1998/Accepted 31 August 1998

Efficient splicing of the 5'-most intron of pre-mRNA requires a 5' m7G(5')ppp(5')N cap, which has been implicated in U1 snRNP binding to 5' splice sites. We demonstrate that the cap alters the kinetic profile of U1 snRNP binding, but its major effect is on U6 snRNA binding. With two alternative wild-type splice sites in an adenovirus pre-mRNA, the cap selectively alters U1 snRNA binding at the site to which cap-independent U1 snRNP binding is stronger and that is used predominantly in splicing; with two consensus sites, the cap acts on both, even though one is substantially preferred for splicing. However, the most striking quantitative effect of the 5' cap is neither on U1 snRNP binding nor on the assembly of large complexes but on the replacement of U1 snRNP by U6 snRNA at the 5' splice site. Inhibition of splicing by a cap analogue is correlated with the loss of U6 interactions at the 5' splice site and not with any loss of U1 snRNP binding.

* Corresponding author. Mailing address: Department of Biochemistry, University of Leicester, Leicester LE1 7RH, United Kingdom. Phone: (116) 2523482. Fax: (116) 2523369. E-mail: eci{at}le.ac.uk.

Molecular and Cellular Biology, December 1998, p. 7510-7520, Vol. 18, No. 12
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Vincent, K., Wang, Q., Jay, S., Hobbs, K., Rymond, B. C. (2003). Genetic Interactions With CLF1 Identify Additional Pre-mRNA Splicing Factors and a Link Between Activators of Yeast Vesicular Transport and Splicing. Genetics 164: 895-907 [Abstract] [Full Text]  
  • Eperon, I. C., Makarova, O. V., Mayeda, A., Munroe, S. H., Cáceres, J. F., Hayward, D. G., Krainer, A. R. (2000). Selection of Alternative 5' Splice Sites: Role of U1 snRNP and Models for the Antagonistic Effects of SF2/ASF and hnRNP A1. Mol. Cell. Biol. 20: 8303-8318 [Abstract] [Full Text]  


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