Detection of antisense protein (ASP) RNA transcripts in individuals infected with human immunodeficiency virus type 1 (HIV-1).

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Version: Final published version
License: CC BY 4.0
Serval ID
serval:BIB_4178D396A6C4
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Detection of antisense protein (ASP) RNA transcripts in individuals infected with human immunodeficiency virus type 1 (HIV-1).
Journal
The Journal of general virology
Author(s)
Mancarella A., Procopio F.A., Achsel T., De Crignis E., Foley B.T., Corradin G., Bagni C., Pantaleo G., Graziosi C.
ISSN
1465-2099 (Electronic)
ISSN-L
0022-1317
Publication state
Published
Issued date
05/2019
Peer-reviewed
Oui
Volume
100
Number
5
Pages
863-876
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
The detection of antisense RNA is hampered by reverse transcription (RT) non-specific priming, due to the ability of RNA secondary structures to prime RT in the absence of specific primers. The detection of antisense RNA by conventional RT-PCR does not allow assessment of the polarity of the initial RNA template, causing the amplification of non-specific cDNAs. In this study we have developed a modified protocol for the detection of human immunodeficiency virus type 1 (HIV-1) antisense protein (ASP) RNA. Using this approach, we have identified ASP transcripts in CD4+ T cells isolated from five HIV-infected individuals, either untreated or under suppressive therapy. We show that ASP RNA can be detected in stimulated CD4+ T cells from both groups of patients, but not in unstimulated cells. We also show that in untreated patients, the patterns of expression of ASP and env are very similar, with the levels of ASP RNA being markedly lower than those of env. Treatment of cells from one viraemic patient with α-amanitin greatly reduces the rate of ASP RNA synthesis, suggesting that it is associated with RNA polymerase II, the central enzyme in the transcription of protein-coding genes. Our data represent the first nucleotide sequences obtained in patients for ASP, demonstrating that its transcription indeed occurs in those HIV-1 lineages in which the ASP open reading frame is present.
Keywords
Adult, Base Sequence/genetics, CD4-Positive T-Lymphocytes/virology, Gene Expression Regulation, Viral/genetics, HIV Infections/virology, HIV-1/genetics, Humans, Male, Middle Aged, Open Reading Frames/genetics, RNA, Antisense/genetics, RNA, Viral/genetics, Virus Replication/genetics, Young Adult, ASP, HIV-1, antisense protein, antisense transcription, envelope
Pubmed
Web of science
Open Access
Yes
Create date
09/04/2019 8:23
Last modification date
28/02/2020 7:19
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