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Home / Equine Research Bank / J Virol / Structural and functional characterization of rev-like trans...
Journal of virology1993; 67(9); 5640-5646; doi: 10.1128/JVI.67.9.5640-5646.1993

Structural and functional characterization of rev-like transcripts of equine infectious anemia virus.

Abstract: Three cDNA clones representing structurally distinct transcripts were isolated from a cDNA library prepared from cells infected with equine infectious anemia virus (EIAV) by using a probe representing the S3 open reading frame, which is thought to encode Rev. One species, designated p2/2, contained four exons and was identical to a previously described polycistronic mRNA that encodes Tat. This transcript was predicted to also direct the synthesis of a truncated form of the transmembrane protein and a putative Rev protein whose N-terminal 29 amino acids, derived from env, are linked to S3 sequences. The second cDNA, p176, also consisted of four exons which were generated by two of three of the same splicing events that occur with p2/2 but not with the Tat mRNA. The alternative splice site giving rise to the second exon of p176 results in a bicistronic message that would encode the same transmembrane and Rev proteins as p2/2. The first exon of the third transcript, p20, was identical to those of p2/2 and p176 but was spliced directly to S3. This monocistronic message could encode a second form of Rev that lacks env sequences, provided that Rev synthesis would initiate at a non-AUG codon. The coding capacity of each cDNA was assessed in a eukaryotic system using S3 antisera. Two putative Rev proteins with apparent molecular masses of 18 and 16 kDa were expressed by p2/2 and p176, while p20 expressed only a 16-kDa species. Analysis of EIAV-infected cells with S3 antisera revealed the presence of an 18-kDa protein. Surprisingly, the same protein was detected in purified virions. By using a reporter construct, the chloramphenicol acetyltransferase gene linked to EIAV env sequences, we were able to demonstrate greatly enhanced chloramphenicol acetyltransferase activity in cells cotransfected with this construct and any of the three cDNAs.
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Publication Date: 1993-09-01 PubMed ID: 8394464PubMed Central: PMC237968DOI: 10.1128/JVI.67.9.5640-5646.1993Google Scholar: Lookup
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  • Non-U.S. Gov't
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  • Non-P.H.S.

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.

This study identifies and characterizes three distinct cDNA clones, a type of artificial DNA, originating from cells infected by the Equine Infectious Anemia Virus (EIAV). Through analyzing each clone, the researchers found two variations of a protein, Rev, which plays a role in viral replication. Surprisingly, one type of Rev was found in the viral particles themselves.

Methods and Techniques

  • The researchers started by isolating three cDNA clones from cells infected with equine infectious anemia virus (EIAV). They used a probe that represented the S3 open reading frame, which is believed to encode the Rev protein.
  • Three cDNA clones were isolated, each representing distinct transcripts. They were named p2/2, p176, and p20.
  • Each of the cDNA clones was studied to ascertain their anatomy and the proteins they could potentially encode.
  • The coding capacity of each cDNA was then tested in a eukaryotic system using S3 antisera. The researchers were able to detect two putative Rev proteins with molecular masses of 18 and 16kDa.

Results and Findings

  • The first cDNA clone, p2/2, was identical to a previously identified mRNA that encodes Tat, a protein crucial for viral replication. This cDNA clone was predicted to direct the synthesis of a truncated form of the transmembrane protein and a Rev protein.
  • The second clone, p176, also generated a protein similar to the Rev protein produced by p2/2 due to similar splicing events.
  • The third cDNA, p20, could encode a second form of Rev without env sequences if Rev synthesis would initiate at a non-AUG codon.
  • By studying EIAV-infected cells with S3 antisera, an 18-kDa protein was detected, surprisingly, this protein was also found in purified virus particles.

Conclusions and Implications

  • The Rev protein plays a significant role in the replication of HIV-type viruses. This study provides the groundwork for a more comprehensive understanding of the various forms of the Rev protein in EIAV and contributes to the broader understanding of the virus’s molecular biology.
  • The presence of a variation of the Rev protein was surprisingly detected in the actual virus particles, a finding that extends our understanding of the role of Rev protein and its variants in the viral life cycle.
  • With these results, the research provides a step toward understanding the complexity of EIAV, which can contribute to the development of effective treatments and vaccines against viral diseases such as HIV.

Cite This Article

APA
Rosin-Arbesfeld R, Rivlin M, Noiman S, Mashiah P, Yaniv A, Miki T, Tronick SR, Gazit A. (1993). Structural and functional characterization of rev-like transcripts of equine infectious anemia virus. J Virol, 67(9), 5640-5646. https://doi.org/10.1128/JVI.67.9.5640-5646.1993

Publication

Journal of virology
ISSN: 0022-538X
NlmUniqueID: 0113724
Country: United States
Language: English
Volume: 67
Issue: 9
Pages: 5640-5646

Researcher Affiliations

Rosin-Arbesfeld, R
  • Department of Human Microbiology, Sackler School of Medicine, Tel Aviv University, Israel.
Rivlin, M
    Noiman, S
      Mashiah, P
        Yaniv, A
          Miki, T
            Tronick, S R
              Gazit, A

                MeSH Terms

                • Amino Acid Sequence
                • Animals
                • Base Sequence
                • Cell Line
                • Chloramphenicol O-Acetyltransferase / metabolism
                • Cloning, Molecular
                • Exons
                • Gene Expression
                • Gene Library
                • Gene Products, rev / biosynthesis
                • Gene Products, rev / genetics
                • Genes, env
                • Genes, rev
                • Genome, Viral
                • Infectious Anemia Virus, Equine / genetics
                • Infectious Anemia Virus, Equine / metabolism
                • Kinetics
                • Molecular Sequence Data
                • Molecular Weight
                • Open Reading Frames
                • Protein Biosynthesis
                • Recombinant Fusion Proteins / metabolism
                • Transcription, Genetic
                • Transfection

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                Citations

                This article has been cited 8 times.
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