Dynamic association of proteins with the pre-mRNA branch region

A. M. MacMillan, Charles C. Query, C. R. Allerson, S. Chen, G. L. Verdine, P. A. Sharp

Research output: Contribution to journalArticle

118 Citations (Scopus)

Abstract

The association of proteins with the branch site region during pre-mRNA splicing was probed using a novel methodology to site-specifically modify the pre-mRNA with the photo-reagent benzophenone. Three sets of proteins were distinguished by the kinetics of their associations with pre-mRNAs, by their association with discrete splicing complexes, and by their differing factor requirements. An early U1 snRNP-dependent cross-link of the branch region to a p80 species was followed by cross-links to p14, p35, and p150 polypeptides associated with the U2 snRNP-pre-mRNA complex. Concomitant with formation of the spliceosome, a rearrangement of protein factors about the branch region occurred, in which the p35 and p150 cross-links were replaced by p220 and p70 species. These results establish that the branch region is recognized in a dynamic fashion by multiple distinct proteins during the course of spliceosomal assembly.

Original languageEnglish (US)
Pages (from-to)3008-3020
Number of pages13
JournalGenes and Development
Volume8
Issue number24
StatePublished - 1994
Externally publishedYes

Fingerprint

RNA Precursors
Proteins
U2 Small Nuclear Ribonucleoproteins
U1 Small Nuclear Ribonucleoproteins
Spliceosomes
Peptides

Keywords

  • branch site
  • photo-cross-linking
  • RNA splicing
  • site-specific modifications

ASJC Scopus subject areas

  • Developmental Biology
  • Genetics

Cite this

MacMillan, A. M., Query, C. C., Allerson, C. R., Chen, S., Verdine, G. L., & Sharp, P. A. (1994). Dynamic association of proteins with the pre-mRNA branch region. Genes and Development, 8(24), 3008-3020.

Dynamic association of proteins with the pre-mRNA branch region. / MacMillan, A. M.; Query, Charles C.; Allerson, C. R.; Chen, S.; Verdine, G. L.; Sharp, P. A.

In: Genes and Development, Vol. 8, No. 24, 1994, p. 3008-3020.

Research output: Contribution to journalArticle

MacMillan, AM, Query, CC, Allerson, CR, Chen, S, Verdine, GL & Sharp, PA 1994, 'Dynamic association of proteins with the pre-mRNA branch region', Genes and Development, vol. 8, no. 24, pp. 3008-3020.
MacMillan AM, Query CC, Allerson CR, Chen S, Verdine GL, Sharp PA. Dynamic association of proteins with the pre-mRNA branch region. Genes and Development. 1994;8(24):3008-3020.
MacMillan, A. M. ; Query, Charles C. ; Allerson, C. R. ; Chen, S. ; Verdine, G. L. ; Sharp, P. A. / Dynamic association of proteins with the pre-mRNA branch region. In: Genes and Development. 1994 ; Vol. 8, No. 24. pp. 3008-3020.
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