Enhancer elements activate the weak 3' splice site of alpha-tropomyosin exon 2. Journal Article uri icon

Overview

abstract

  • We have identified four purine-rich sequences that act as splicing enhancer elements to activate the weak 3' splice site of alpha-tropomyosin exon 2. These elements also activate the splicing of heterologous substrates containing weak 3' splice sites or mutated 5' splice sites. However, they are unique in that they can activate splicing whether they are placed in an upstream or downstream exon, and the two central elements can function regardless of their position relative to one another. The presence of excess RNAs containing these enhancers could effectively inhibit in vitro pre-mRNA splicing reactions in a substrate-dependent manner and, at lower concentrations of competitor RNA, the addition of SR proteins could relieve the inhibition. However, when extracts were depleted by incubation with biotinylated exon 2 RNAs followed by passage over streptavidin agarose, SR proteins were not sufficient to restore splicing. Instead, both SR proteins and fractions containing a 110-kD protein were necessary to rescue splicing. Using gel mobility shift assays, we show that formation of stable enhancer-specific complexes on alpha-tropomyosin exon 2 requires the presence of both SR proteins and the 110-kD protein. By analogy to the doublesex exon enhancer elements in Drosophila, our results suggest that assembly of mammalian exon enhancer complexes requires both SR and non-SR proteins to activate selection of weak splice sites.

publication date

  • December 1, 1998

has subject area

Date in CU Experts

  • March 13, 2015 1:10 AM

Full Author List

  • Dye BT; Buvoli M; Mayer SA; Lin CH; Patton JG

author count

  • 5

citation count

  • 17

published in

  • RNA  Journal

Other Profiles

International Standard Serial Number (ISSN)

  • 1355-8382

Electronic International Standard Serial Number (EISSN)

  • 1469-9001

Additional Document Info

start page

  • 1523

end page

  • 1536

volume

  • 4

issue

  • 12