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

I. C. Eperon, D. C. Ireland, R. A. Smith, A. Mayeda, A. R. Krainer

Research output: Contribution to journalArticle

153 Citations (Scopus)

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 snRNrPs 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.

Original languageEnglish
Pages (from-to)3607-3617
Number of pages11
JournalEMBO Journal
Volume12
Issue number9
Publication statusPublished - 01-01-1993

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U1 Small Nuclear Ribonucleoproteins
Small Nuclear Ribonucleoproteins
RNA Splice Sites
Ribonuclease H
RNA Precursors
Genes
Molecules
Adenoviridae

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Eperon, I. C., Ireland, D. C., Smith, R. A., Mayeda, A., & Krainer, A. R. (1993). Pathways for selection of 5' splice sites by U1 snRNPs and SF2/ASF. EMBO Journal, 12(9), 3607-3617.
Eperon, I. C. ; Ireland, D. C. ; Smith, R. A. ; Mayeda, A. ; Krainer, A. R. / Pathways for selection of 5' splice sites by U1 snRNPs and SF2/ASF. In: EMBO Journal. 1993 ; Vol. 12, No. 9. pp. 3607-3617.
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Eperon, IC, Ireland, DC, Smith, RA, Mayeda, A & Krainer, AR 1993, 'Pathways for selection of 5' splice sites by U1 snRNPs and SF2/ASF', EMBO Journal, vol. 12, no. 9, pp. 3607-3617.

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.

In: EMBO Journal, Vol. 12, No. 9, 01.01.1993, p. 3607-3617.

Research output: Contribution to journalArticle

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Eperon IC, Ireland DC, Smith RA, Mayeda A, Krainer AR. Pathways for selection of 5' splice sites by U1 snRNPs and SF2/ASF. EMBO Journal. 1993 Jan 1;12(9):3607-3617.