Pathways for selection of 5' splice sites by U1 snRNPs and SF2/ASF

Eperon, I. C., Ireland, D. C., Smith, R. A., Mayeda, A., Krainer, A. R. (September 1993) Pathways for selection of 5' splice sites by U1 snRNPs and SF2/ASF. EMBO Journal, 12 (9). pp. 3607-3617. ISSN 0261-4189

URL: http://www.ncbi.nlm.nih.gov/pubmed/8253084

Abstract

We have used protection against ribonuclease H to investigate the mechanisms by which U1 small nuclear ribonucleoprotein particles (snRNPs) determine the use of two alternative 5' splice sites. The initial binding of U1 snRNPs to alternative consensus splice sites was indiscriminate, and on a high proportion of pre-mRNA molecules both sites were occupied simultaneously. When the sites were close, this inhibited splicing. We propose that double occupancy leads to the use of the downstream site for splicing and that this is the cause of the proximity effect seen with strong alternative splice sites. This model predicts that splicing to an upstream site of any strength requires a low affinity of U1 snRNPs for the downstream site. This prediction was tested both by cleaving the 5' end of U1 snRNA and by altering the sequence of the downstream site of an adenovirus E1A gene. The enhancement of downstream 5' splice site use by splicing factor SF2/ASF appears to be mediated by an increase in the strength of U1 snRNP binding to all sites indiscriminately.

Item Type: Paper
Uncontrolled Keywords: asf ribonuclease-h protection sf2 splice site selection u1 snrnps pre-messenger-rna small nuclear ribonucleoprotein junction sequences mammalian nuclei conserved family alternative 5' invitro invivo proteins u2
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > nuclear ribonucleoprotein
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > pre-mRNA
CSHL Authors:
Communities: CSHL labs > Krainer lab
Depositing User: Matt Covey
Date: September 1993
Date Deposited: 11 Dec 2013 14:52
Last Modified: 11 Dec 2013 14:52
PMCID: PMC413636
URI: http://repository.cshl.edu/id/eprint/28952

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