Mutations altering acetylated residues in the CTD of HIV-1 integrase cause defects in proviral transcription at early times after integration of viral DNA

• Published in: PLoS pathogens Vol. 16; no. 12; p. e1009147
• Main Authors: Winans, Shelby, Goff, Stephen P.
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 22.12.2020; Public Library of Science (PLoS)
• Subjects: Acetylation; Analysis; Binding sites; Biology and Life Sciences; Cell Line; Chromatin; Circuits; Deoxyribonucleic acid; Diagnosis; DNA; DNA methylation; DNA, Viral - physiology; DNA-directed RNA polymerase; Gene expression; Gene mutations; Gene therapy; Genetic aspects; Genetic transcription; Genomes; HIV; HIV infection; HIV Integrase - genetics; HIV-1 - genetics; Human immunodeficiency virus; Humans; Infections; Integrase; Integration; Medicine and Health Sciences; Mutation; Phosphorylation; Physical Sciences; Protein Processing, Post-Translational - genetics; Proteins; Proviruses - genetics; Retrovirus infections; Ribonucleic acid; RNA; RNA polymerase; Site selection; Tat protein; Transcription factors; Transcription initiation; Viral Transcription - genetics; Virus Integration - genetics; United States
• ISSN: 1553-7374; 1553-7366; 1553-7374
• DOI: 10.1371/journal.ppat.1009147

The central function of the retroviral integrase protein (IN) is to catalyze the integration of viral DNA into the host genome to form the provirus. The IN protein has also been reported to play a role in a number of other processes throughout the retroviral life cycle such as reverse transcription, nuclear import and particle morphogenesis. Studies have shown that HIV-1 IN is subject to multiple post-translational modifications (PTMs) including acetylation, phosphorylation and SUMOylation. However, the importance of these modifications during infection has been contentious. In this study we attempt to clarify the role of acetylation of HIV-1 IN during the retroviral life cycle. We show that conservative mutation of the known acetylated lysine residues has only a modest effect on reverse transcription and proviral integration efficiency in vivo . However, we observe a large defect in successful expression of proviral genes at early times after infection by an acetylation-deficient IN mutant that cannot be explained by delayed integration dynamics. We demonstrate that the difference between the expression of proviruses integrated by an acetylation mutant and WT IN is likely not due to altered integration site distribution but rather directly due to a lower rate of transcription. Further, the effect of the IN mutation on proviral gene expression is independent of the Tat protein or the LTR promoter. At early times after integration when the transcription defect is observed, the LTRs of proviruses integrated by the mutant IN have altered histone modifications as well as reduced IN protein occupancy. Over time as the transcription defect in the mutant virus diminishes, histone modifications on the WT and mutant proviral LTRs reach comparable levels. These results highlight an unexpected role for the IN protein in regulating proviral transcription at early times post-integration.