Abstract | UNLABELLED: Approximately 8% of the human genome is made up of endogenous retroviral sequences. As the HIV-1 Tat protein activates the overall expression of the human endogenous retrovirus type K (HERV-K) (HML-2), we used next-generation sequencing to determine which of the 91 currently annotated HERV-K (HML-2) proviruses are regulated by Tat. Transcriptome sequencing of total RNA isolated from Tat- and vehicle-treated peripheral blood lymphocytes from a healthy donor showed that Tat significantly activates expression of 26 unique HERV-K (HML-2) proviruses, silences 12, and does not significantly alter the expression of the remaining proviruses. Quantitative reverse transcription-PCR validation of the sequencing data was performed on Tat-treated PBLs of seven donors using provirus-specific primers and corroborated the results with a substantial degree of quantitative similarity. IMPORTANCE: The expression of HERV-K (HML-2) is tightly regulated but becomes markedly increased following infection with HIV-1, in part due to the HIV-1 Tat protein. The findings reported here demonstrate the complexity of the genome-wide regulation of HERV-K (HML-2) expression by Tat. This work also demonstrates that although HERV-K (HML-2) proviruses in the human genome are highly similar in terms of DNA sequence, modulation of the expression of specific proviruses in a given biological situation can be ascertained using next-generation sequencing and bioinformatics analysis.
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Authors | Marta J Gonzalez-Hernandez, James D Cavalcoli, Maureen A Sartor, Rafael Contreras-Galindo, Fan Meng, Manhong Dai, Derek Dube, Anjan K Saha, Scott D Gitlin, Gilbert S Omenn, Mark H Kaplan, David M Markovitz |
Journal | Journal of virology
(J Virol)
Vol. 88
Issue 16
Pg. 8924-35
(Aug 2014)
ISSN: 1098-5514 [Electronic] United States |
PMID | 24872592
(Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
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Copyright | Copyright © 2014, American Society for Microbiology. All Rights Reserved. |
Chemical References |
- Gene Products, tat
- RNA, Viral
- Viral Proteins
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Topics |
- Cells, Cultured
- Endogenous Retroviruses
(genetics, metabolism)
- Gene Products, tat
(genetics, metabolism)
- Genome, Human
(genetics)
- HIV Infections
(genetics, metabolism)
- HIV-1
(genetics, metabolism)
- High-Throughput Nucleotide Sequencing
(methods)
- Humans
- Lymphocytes
(virology)
- Proviruses
(genetics, metabolism)
- RNA, Viral
(genetics)
- Transcriptome
(genetics)
- Viral Proteins
(genetics, metabolism)
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