Efficient polyadenylation of Rous sarcoma virus RNA requires the negative regulator of splicing element. Nucleic Acids Res 2002 Feb 01;30(3):810-7
Date
01/26/2002Pubmed ID
11809895Pubmed Central ID
PMC100303DOI
10.1093/nar/30.3.810Scopus ID
2-s2.0-0036464505 (requires institutional sign-in at Scopus site) 25 CitationsAbstract
Rous sarcoma virus pre-mRNA contains an element known as the negative regulator of splicing (NRS) that acts to inhibit viral RNA splicing. The NRS binds serine/arginine-rich (SR) proteins, hnRNP H and the U1/U11 snRNPs, and appears to inhibit splicing by acting as a decoy 5' splice site. Deletions within the gag gene that encompass the NRS also lead to increased read-through past the viral polyadenylation site, suggesting a role for the NRS in promoting polyadenylation. Using NRS-specific deletions and mutations, we show here that a polyadenylation stimulatory activity maps directly to the NRS and is most likely dependent upon SR proteins and U1 and/or U11 snRNP. hnRNP H does not appear to mediate splicing control or stimulate RSV polyadenylation, since viral RNAs containing hnRNP H-specific mutations were spliced and polyadenylated normally. However, the ability of hnRNP H mutations to suppress the read-through caused by an SR protein mutation suggests the potential for hnRNP H to antagonize polyadenylation. Interestingly, disruption of splicing control closely correlated with increased read-through, indicating that a functional NRS is necessary for efficient RSV polyadenylation rather than binding of an individual factor. We propose a model in which the NRS serves to enhance polyadenylation of RSV unspliced RNA in a process analogous to the stimulation of cellular pre-mRNA polyadenylation by splicing complexes.
Author List
Fogel BL, McNally LM, McNally MTMESH terms used to index this publication - Major topics in bold
AnimalsAvian Sarcoma Viruses
Binding Sites
Chick Embryo
Fibroblasts
Genes, Viral
HeLa Cells
Heterogeneous-Nuclear Ribonucleoprotein Group F-H
Heterogeneous-Nuclear Ribonucleoproteins
Humans
Models, Genetic
Mutation
Nuclear Proteins
Nuclease Protection Assays
Phosphoproteins
Polyadenylation
Protein Binding
RNA Precursors
RNA Splicing
RNA, Viral
RNA-Binding Proteins
Regulatory Sequences, Nucleic Acid
Ribonucleoprotein, U1 Small Nuclear
Ribonucleoproteins
Serine-Arginine Splicing Factors
Two-Hybrid System Techniques
