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Molecular and cellular biology2000; 20(10); 3550-3557; doi: 10.1128/MCB.20.10.3550-3557.2000

Binding of equine infectious anemia virus rev to an exon splicing enhancer mediates alternative splicing and nuclear export of viral mRNAs.

Abstract: In addition to facilitating the nuclear export of incompletely spliced viral mRNAs, equine infectious anemia virus (EIAV) Rev regulates alternative splicing of the third exon of the tat/rev mRNA. In the presence of Rev, this exon of the bicistronic RNA is skipped in a fraction of the spliced mRNAs. In this report, the cis-acting requirements for exon 3 usage were correlated with sequences necessary for Rev binding and transport of incompletely spliced RNA. The presence of a purine-rich exon splicing enhancer (ESE) was required for exon 3 recognition, and the addition of Rev inhibited exon 3 splicing. Glutathione-S-transferase (GST)-Rev bound to probes containing the ESE, and mutation of GAA repeats to GCA within the ESE inhibited both exon 3 recognition in RNA splicing experiments and GST-Rev binding in vitro. These results suggest that Rev regulates alternative splicing by binding at or near the ESE to block SR protein-ESE interactions. A 57-nucleotide sequence containing the ESE was sufficient to mediate Rev-dependent nuclear export of incompletely spliced RNAs. Rev export activity was significantly inhibited by mutation of the ESE or by trans-complementation with SF2/ASF. These results indicate that the ESE functions as a Rev-responsive element and demonstrate that EIAV Rev mediates exon 3 exclusion through protein-RNA interactions required for efficient export of incompletely spliced viral RNAs.
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Publication Date: 2000-04-25 PubMed ID: 10779344PubMed Central: PMC85647DOI: 10.1128/MCB.20.10.3550-3557.2000Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article explores how equine infectious anemia virus (EIAV) Rev protein influences the splicing and nuclear export of viral mRNAs. The findings reveal that the Rev protein binds to a specific sequence in the RNA to regulate alternative splicing and enables the effective exit of incomplete viral RNA from the cell nucleus.

Understanding Rev’s Role in EIAV

  • The equine infectious anemia virus (EIAV) Rev protein does more than just facilitate the nuclear export of incompletely spliced viral mRNAs. It also regulates the splicing of the third exon in the tat/rev mRNA—part of the instructions for producing viral proteins.
  • When the Rev protein is present, some spliced mRNAs skip this third exon in the bicistronic RNA, a type of RNA that can be read in two different ways to produce different products.
  • The researchers correlated the requirements for using this third exon with the necessary sequences for the binding of Rev and the transportation of incomplete RNA.

Functions of the Exon Splicing Enhancer

  • The presence of a purine-rich exon splicing enhancer (ESE), a sequence that promotes the recognition of an exon during the splicing process, is required for the third exon’s recognition.
  • When Rev was added, the researchers found that it inhibited the splicing of this exon. They observed that under certain conditions, the Glutathione-S-transferase (GST)-Rev protein bound to probes that contained this ESE.
  • Experiments revealed that changes to the sequence of this ESE, specifically from GAA repeats to GCA, barred both the recognition of the third exon in RNA splicing experiments and binding to the GST-Rev protein.

Rev’s Mediating Activity and the ESE

  • These results led the researchers to posit that Rev might regulate alternative splicing by binding at or near the ESE, hence blocking the interaction between other proteins and the ESE.
  • A 57-nucleotide long sequence containing the ESE was found to adequately mediate the export of incomplete RNAs from the nucleus depending on the presence of Rev.
  • They discovered that mutations in the ESE or adding in an additional external protein (SF2/ASF) in trans-complementation significantly impaired the function of the Rev protein in exporting RNA.

Implications of Findings

  • The ESE’s functions indicate that it acts as a responsive element to Rev, indicating a crucial role in the virus’s mechanism.
  • The EIAV Rev protein manipulates exon 3 exclusion via protein-RNA interactions which are needed for the efficient exportation of incompletely spliced viral RNAs.
  • This information could be significant in devising treatments for diseases caused by the EIAV, as disrupting this interaction might affect the virus’s ability to replicate.

Cite This Article

APA
Belshan M, Park GS, Bilodeau P, Stoltzfus CM, Carpenter S. (2000). Binding of equine infectious anemia virus rev to an exon splicing enhancer mediates alternative splicing and nuclear export of viral mRNAs. Mol Cell Biol, 20(10), 3550-3557. https://doi.org/10.1128/MCB.20.10.3550-3557.2000

Publication

Molecular and cellular biology
ISSN: 0270-7306
NlmUniqueID: 8109087
Country: United States
Language: English
Volume: 20
Issue: 10
Pages: 3550-3557

Researcher Affiliations

Belshan, M
  • Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, Iowa 50011, USA.
Park, G S
    Bilodeau, P
      Stoltzfus, C M
        Carpenter, S

          MeSH Terms

          • Alternative Splicing
          • Binding Sites
          • Biological Transport
          • Cell Nucleus
          • Exons / genetics
          • Gene Products, rev / genetics
          • Gene Products, rev / metabolism
          • Infectious Anemia Virus, Equine / genetics
          • Mutagenesis
          • Nuclear Proteins / metabolism
          • Protein Binding
          • RNA, Messenger / genetics
          • RNA, Messenger / metabolism
          • RNA, Viral / genetics
          • RNA, Viral / metabolism
          • RNA-Binding Proteins
          • Serine-Arginine Splicing Factors

          Grant Funding

          • R01 AI036073 / NIAID NIH HHS
          • R56 AI036073 / NIAID NIH HHS
          • AI36073 / NIAID NIH HHS
          • CA 28951 / NCI NIH HHS

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          This article has been cited 16 times.
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