Structural and Functional Analysis of the Rous Sarcoma Virus Negative Regulator of Splicing and Demonstration of Its Activation by the 9G8 SR Protein

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2010 Dec 25

PMID 21183462

Citations 1

Authors

Aileen Bar

Virginie Marchand

Georges Khoury

Natacha Dreumont

Annie Mougin

Nathalie Robas

James Stevenin

Athanase Visvikis

Christiane Branlant

Affiliations

Soon will be listed here.

Abstract

Retroviruses require both spliced and unspliced RNAs for replication. Accumulation of Rous Sarcoma virus (RSV) unspliced RNA depends upon the negative regulator of splicing (NRS). Its 5'-part is considered as an ESE binding SR proteins. Its 3'-part contains a decoy 5'-splice site (ss), which inhibits splicing at the bona fide 5'-ss. Only the 3D structure of a small NRS fragment had been experimentally studied. Here, by chemical and enzymatic probing, we determine the 2D structure of the entire RSV NRS. Structural analysis of other avian NRSs and comparison with all sequenced avian NRSs is in favour of a phylogenetic conservation of the NRS 2D structure. By combination of approaches: (i) in vitro and in cellulo splicing assays, (ii) footprinting assays and (iii) purification and analysis of reconstituted RNP complex, we define a small NRS element retaining splicing inhibitory property. We also demonstrate the capability of the SR protein 9G8 to increase NRS activity in vitro and in cellulo. Altogether these data bring new insights on how NRS fine tune splicing activity.

Citing Articles

The RNA-Binding Proteins SRP14 and HMGB3 Control HIV-1 Tat mRNA Processing and Translation During HIV-1 Latency.

Khoury G, Lee M, Ramarathinam S, McMahon J, Purcell A, Sonza S Front Genet. 2021; 12:680725.

PMID: 34194479 PMC: 8236859. DOI: 10.3389/fgene.2021.680725.

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