title: Yeast genetic analysis reveals the involvement of chromatin reassembly factors in repressing HIV-1 basal transcription.
abstract: Rebound of HIV viremia after interruption of anti-retroviral therapy is due to the small population of CD4+ T cells that remain latently infected. HIV-1 transcription is the main process controlling post-integration latency. Regulation of HIV-1 transcription takes place at both initiation and elongation levels. Pausing of RNA polymerase II at the 5' end of HIV-1 transcribed region (5'HIV-TR), which is immediately downstream of the transcription start site, plays an important role in the regulation of viral expression. The activation of HIV-1 transcription correlates with the rearrangement of a positioned nucleosome located at this region. These two facts suggest that the 5'HIV-TR contributes to inhibit basal transcription of those HIV-1 proviruses that remain latently inactive. However, little is known about the cell elements mediating the repressive role of the 5'HIV-TR. We performed a genetic analysis of this phenomenon in Saccharomyces cerevisiae after reconstructing a minimal HIV-1 transcriptional system in this yeast. Unexpectedly, we found that the critical role played by the 5'HIV-TR in maintaining low levels of basal transcription in yeast is mediated by FACT, Spt6, and Chd1, proteins so far associated with chromatin assembly and disassembly during ongoing transcription. We confirmed that this group of factors plays a role in HIV-1 postintegration latency in human cells by depleting the corresponding human orthologs with shRNAs, both in HIV latently infected cell populations and in particular single-integration clones, including a latent clone with a provirus integrated in a highly transcribed gene. Our results indicate that chromatin reassembly factors participate in the establishment of the equilibrium between activation and repression of HIV-1 when it integrates into the human genome, and they open the possibility of considering these factors as therapeutic targets of HIV-1 latency.
authors.affiliation: Departamento de Genética, Universidad de Sevilla, Seville, Spain.
authors.forename: Manuela
authors.initials: M
authors.lastname: Vanti
authors.forename: Edurne
authors.initials: E
authors.lastname: Gallastegui
authors.forename: Iñaki
authors.initials: I
authors.lastname: Respaldiza
authors.forename: Alfonso
authors.initials: A
authors.lastname: Rodríguez-Gil
authors.forename: Fernando
authors.initials: F
authors.lastname: Gómez-Herreros
authors.forename: Silvia
authors.initials: S
authors.lastname: Jimeno-González
authors.forename: Albert
authors.initials: A
authors.lastname: Jordan
authors.forename: Sebastián
authors.initials: S
authors.lastname: Chávez
mesh_terms: D002843:Chromatin / Q000378:metabolism*; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D008954:Models, Biological; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; C100927:SUPT6H protein, human; D014157:Transcription Factors; D012319:RNA Polymerase II; D004265:DNA Helicases; C080359:CHD1 protein, human