Article: article from journal or magazin.
Analysis of HIV-1 expression level and sense of transcription by high-throughput sequencing of the infected cell.
Journal of Virology
Publication types: Journal Article ; Research Support, Non-U.S. Gov't Publication Status: ppublish
Next-generation sequencing offers an unprecedented opportunity to jointly analyze cellular and viral transcriptional activity without prerequisite knowledge of the nature of the transcripts. SupT1 cells were infected with a vesicular stomatitis virus G envelope protein (VSV-G)-pseudotyped HIV vector. At 24 h postinfection, both cellular and viral transcriptomes were analyzed by serial analysis of gene expression followed by high-throughput sequencing (SAGE-Seq). Read mapping resulted in 33 to 44 million tags aligning with the human transcriptome and 0.23 to 0.25 million tags aligning with the genome of the HIV-1 vector. Thus, at peak infection, 1 transcript in 143 is of viral origin (0.7%), including a small component of antisense viral transcription. Of the detected cellular transcripts, 826 (2.3%) were differentially expressed between mock- and HIV-infected samples. The approach also assessed whether HIV-1 infection modulates the expression of repetitive elements or endogenous retroviruses. We observed very active transcription of these elements, with 1 transcript in 237 being of such origin, corresponding on average to 123,123 reads in mock-infected samples (0.40%) and 129,149 reads in HIV-1-infected samples (0.45%) mapping to the genomic Repbase repository. This analysis highlights key details in the generation and interpretation of high-throughput data in the setting of HIV-1 cellular infection.
Cell Line, Gene Expression Profiling, Genetic Vectors/genetics, HIV Infections/virology, HIV-1/genetics, HIV-1/metabolism, High-Throughput Nucleotide Sequencing/methods, Humans, Membrane Glycoproteins/genetics, Membrane Glycoproteins/metabolism, RNA, Messenger/genetics, RNA, Messenger/metabolism, RNA, Viral/genetics, RNA, Viral/metabolism, Sequence Tagged Sites, T-Lymphocytes/metabolism, T-Lymphocytes/virology, Transcription, Genetic, Viral Envelope Proteins/genetics, Viral Envelope Proteins/metabolism, Virus Replication
Web of science
Last modification date