Dynamic association of proteins with the pre-mRNA branch region

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

Research output: Contribution to journalArticle

123 Scopus citations

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 - Dec 1 1994
Externally publishedYes

Keywords

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

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology

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  • 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.