Faculty Collaboration Database

Mutation of an RSV intronic element abolishes both U11/U12 snRNP binding and negative regulation of splicing. Genes Dev 1993 Oct;7(10):1926-36

Date

10/01/1993

Pubmed ID

8405999

DOI

10.1101/gad.7.10.1926

Scopus ID

2-s2.0-0027501973 (requires institutional sign-in at Scopus site)   77 Citations

Abstract

A cis-acting negative regulator of splicing (NRS) within the gag gene of RSV is involved in control of the relative levels of spliced and unspliced viral mRNAs. Insertion of the NRS into the intron of an adenovirus pre-mRNA resulted in inhibition of splicing in vitro before the first cleavage step. Analyses of spliceosome assembly with this substrate showed that it formed large RNP complexes that did not migrate like mature spliceosomes on native gels. Affinity selection of the RNP complexes formed on NRS-containing pre-mRNAs showed an association with U11 and U12 snRNPs, as well as with the spliceosomal snRNPs. Immunoprecipitation with antisera specific for U1 and U2 snRNPS showed binding of both snRNPs to NRS RNA. A 7-nucleotide missense mutation in the NRS that prevented binding of U11 and U12 snRNPs impaired NRS activity in vivo, suggesting a functional role for U11 and U12 snRNPs in the inhibition of splicing mediated by the RSV NRS RNA.

Author List

Gontarek RR, McNally MT, Beemon K

MESH terms used to index this publication - Major topics in bold

Adenoviridae
Avian Sarcoma Viruses
Base Sequence
Gene Expression Regulation, Viral
Genes, gag
HeLa Cells
Humans
Introns
Macromolecular Substances
Models, Genetic
Molecular Sequence Data
Mutation
RNA Precursors
RNA Splicing
Regulatory Sequences, Nucleic Acid
Ribonucleoprotein, U1 Small Nuclear
Ribonucleoprotein, U2 Small Nuclear
Ribonucleoproteins, Small Nuclear
Spliceosomes