NFAT5 Regulates HIV-1 in Primary Monocytes via a Highly Conserved Long Terminal Repeat Site

PLoS Pathogens

James V Falvo¹  Shahin Ranjbar¹  Sang-Kyung Lee¹  Premlata Shankar¹  Judy Lieberman²  Anne E Goldfeld³  Alla V Tsytsykova⁴  Ricardo Rajsbaum⁴ 

[1] CBR Institute for Biomedical Research, Harvard Medical School, Boston, Massachusetts, United States of America;Department of Bioengineering, Hanyang University, Seoul, Korea;Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America;Department of Pediatrics, Harvard Medical School, Boston, Massachusetts, United States of America

关键词: HIV-1;    Small interfering RNAs;    Transcription factors;    Sequence motif analysis;    Viral replication;    DNA-binding proteins;    DNA transcription;    Macrophages;   

DOI  :  10.1371/journal.ppat.0020130

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

To replicate, HIV-1 capitalizes on endogenous cellular activation pathways resulting in recruitment of key host transcription factors to its viral enhancer. RNA interference has been a powerful tool for blocking key checkpoints in HIV-1 entry into cells. Here we apply RNA interference to HIV-1 transcription in primary macrophages, a major reservoir of the virus, and specifically target the transcription factor NFAT5 (nuclear factor of activated T cells 5), which is the most evolutionarily divergent NFAT protein. By molecularly cloning and sequencing isolates from multiple viral subtypes, and performing DNase I footprinting, electrophoretic mobility shift, and promoter mutagenesis transfection assays, we demonstrate that NFAT5 functionally interacts with a specific enhancer binding site conserved in HIV-1, HIV-2, and multiple simian immunodeficiency viruses. Using small interfering RNA to ablate expression of endogenous NFAT5 protein, we show that the replication of three major HIV-1 viral subtypes (B, C, and E) is dependent upon NFAT5 in human primary differentiated macrophages. Our results define a novel host factor–viral enhancer interaction that reveals a new regulatory role for NFAT5 and defines a functional DNA motif conserved across HIV-1 subtypes and representative simian immunodeficiency viruses. Inhibition of the NFAT5–LTR interaction may thus present a novel therapeutic target to suppress HIV-1 replication and progression of AIDS.

【 授权许可】

CC BY