title: Tight interplay among SAMHD1 protein level, cellular dNTP levels, and HIV-1 proviral DNA synthesis kinetics in human primary monocyte-derived macrophages.
abstract: Recently, SAMHD1 has come under intense focus as a host anti-HIV factor. SAMHD1 is a dNTP triphosphohydrolase, which leads to the regulation of DNA metabolism in host cells. HIV-2/SIV (simian immunodeficiency virus) viral protein x (Vpx) has been shown to promote the degradation of SAMHD1. In this study, we examine the kinetics of SAMHD1 degradation, the increase in the dNTP pool level, and the efficiency of proviral DNA synthesis in Vpx+ virus-like particle (VLP)-treated monocyte-derived macrophages (MDMs). Our results indicate a very close temporal link with a reduction in SAMHD1 detected within the first few hours of Vpx+ VLP treatment. This loss of SAMHD1 is followed by a significant increase in cellular dNTP levels by 8 h after Vpx+ VLP addition, ultimately leading to the enhancement of the HIV proviral DNA synthesis rate and HIV infection in MDMs. Finally, the pretreatment of MDMs with the Vpx+ VLPs, which is a widely used protocol, displayed identical proviral DNA synthesis as compared with MDMs co-treated with Vpx+ VLP and HIV vector. These findings further indicate that Vpx degradation of SAMHD1 is sufficiently rapid to enable appropriate progression of reverse transcription in MDMs, even when present at the time of infection. Overall, this study demonstrates a tight interplay between SAMHD1 level, dNTP levels, and HIV proviral DNA synthesis kinetics in MDMs.
authors.affiliation: Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York 14642, USA. baek_kim@urmc.rochester.edu
authors.forename: Baek
authors.initials: B
authors.lastname: Kim
authors.forename: Laura A
authors.initials: LA
authors.lastname: Nguyen
authors.forename: Waaqo
authors.initials: W
authors.lastname: Daddacha
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Hollenbaugh
mesh_terms: D004261:DNA Replication; D004279:DNA, Viral / Q000378:metabolism*; D015226:Dinucleoside Phosphates / Q000737:chemistry / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D018950:Lipopolysaccharide Receptors / Q000096:biosynthesis; D008264:Macrophages / Q000821:virology*; D008954:Models, Biological; D020559:Monomeric GTP-Binding Proteins / Q000378:metabolism*; D009711:Nucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods; D000076106:SAM Domain and HD Domain-Containing Protein 1; D013997:Time Factors; D054334:Viral Regulatory and Accessory Proteins / Q000378:metabolism*
chemical_list: D004279:DNA, Viral; D015226:Dinucleoside Phosphates; D018950:Lipopolysaccharide Receptors; D009711:Nucleotides; C513059:VPX protein, Simian immunodeficiency virus; D054334:Viral Regulatory and Accessory Proteins; D049452:Green Fluorescent Proteins; D000076106:SAM Domain and HD Domain-Containing Protein 1; C484280:SAMHD1 protein, human; D020559:Monomeric GTP-Binding Proteins