dNTP pool modulation dynamics by SAMHD1 protein in monocyte-derived macrophages

Joseph A. Hollenbaugh, Sijia Tao, Gina M. Lenzi, Sulryung Ryu, Dong Hyun Kim, Felipe Diaz-Griffero, Raymond F. Schinazi, Baek Kim

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Background: SAMHD1 degrades deoxyribonucleotides (dNTPs), suppressing viral DNA synthesis in macrophages. Recently, viral protein X (Vpx) of HIV-2/SIVsm was shown to target SAMHD1 for proteosomal degradation and led to elevation of dNTP levels, which in turn accelerated proviral DNA synthesis of lentiviruses in macrophages.Results: We investigated both time-dependent and quantitative interplays between SAMHD1 level and dNTP concentrations during multiple exposures of Vpx in macrophages. The following were observed. First, SAMHD1 level was rapidly reduced by Vpx + VLP to undetectable levels by Western blot analysis. Recovery of SAMHD1 was very slow with less than 3% of the normal macrophage level detected at day 6 post Vpx treatment and only ~30% recovered at day 14. Second, dGTP, dCTP and dTTP levels peaked at day 1 post Vpx treatment, whereas dATP peaked at day 2. However, all dNTPs rapidly decreased starting at day 3, while SAMHD1 level was below the level of detection. Third, when Vpx pretreated macrophages were re-exposed to a second Vpx treatment at day 7, we observed dNTP elevation that had faster kinetics than the first Vpx + VLP treatment. Moreover, we performed a short kinetic analysis of the second Vpx treatment to find that dATP and dGTP levels peaked at 8 hours post secondary VLP treatment. dGTP peak was consistently higher than the primary, whereas peak dATP concentration was basically equivalent to the first Vpx + VLP treatment. Lastly, HIV-1 replication kinetics were faster in macrophages treated after the secondary Vpx treatments when compared to the initial single Vpx treatment.Conclusion: This study reveals that a very low level of SAMHD1 sufficiently modulates the normally low dNTP levels in macrophages and proposes potential diverse mechanisms of Vpx-mediated dNTP regulation in macrophages.

Original languageEnglish (US)
Article number63
JournalRetrovirology
Volume11
Issue number1
DOIs
StatePublished - Aug 27 2014

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Viral Proteins
Macrophages
Proteins
Deoxyribonucleotides
Lentivirus
Viral DNA
HIV-1
Western Blotting

Keywords

  • dNTPs: virus-like particles
  • HIV-1
  • Monocytes-derived macrophages
  • SAMHD1
  • Vpx

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

dNTP pool modulation dynamics by SAMHD1 protein in monocyte-derived macrophages. / Hollenbaugh, Joseph A.; Tao, Sijia; Lenzi, Gina M.; Ryu, Sulryung; Kim, Dong Hyun; Diaz-Griffero, Felipe; Schinazi, Raymond F.; Kim, Baek.

In: Retrovirology, Vol. 11, No. 1, 63, 27.08.2014.

Research output: Contribution to journalArticle

Hollenbaugh, JA, Tao, S, Lenzi, GM, Ryu, S, Kim, DH, Diaz-Griffero, F, Schinazi, RF & Kim, B 2014, 'dNTP pool modulation dynamics by SAMHD1 protein in monocyte-derived macrophages', Retrovirology, vol. 11, no. 1, 63. https://doi.org/10.1186/s12977-014-0063-2
Hollenbaugh, Joseph A. ; Tao, Sijia ; Lenzi, Gina M. ; Ryu, Sulryung ; Kim, Dong Hyun ; Diaz-Griffero, Felipe ; Schinazi, Raymond F. ; Kim, Baek. / dNTP pool modulation dynamics by SAMHD1 protein in monocyte-derived macrophages. In: Retrovirology. 2014 ; Vol. 11, No. 1.
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abstract = "Background: SAMHD1 degrades deoxyribonucleotides (dNTPs), suppressing viral DNA synthesis in macrophages. Recently, viral protein X (Vpx) of HIV-2/SIVsm was shown to target SAMHD1 for proteosomal degradation and led to elevation of dNTP levels, which in turn accelerated proviral DNA synthesis of lentiviruses in macrophages.Results: We investigated both time-dependent and quantitative interplays between SAMHD1 level and dNTP concentrations during multiple exposures of Vpx in macrophages. The following were observed. First, SAMHD1 level was rapidly reduced by Vpx + VLP to undetectable levels by Western blot analysis. Recovery of SAMHD1 was very slow with less than 3{\%} of the normal macrophage level detected at day 6 post Vpx treatment and only ~30{\%} recovered at day 14. Second, dGTP, dCTP and dTTP levels peaked at day 1 post Vpx treatment, whereas dATP peaked at day 2. However, all dNTPs rapidly decreased starting at day 3, while SAMHD1 level was below the level of detection. Third, when Vpx pretreated macrophages were re-exposed to a second Vpx treatment at day 7, we observed dNTP elevation that had faster kinetics than the first Vpx + VLP treatment. Moreover, we performed a short kinetic analysis of the second Vpx treatment to find that dATP and dGTP levels peaked at 8 hours post secondary VLP treatment. dGTP peak was consistently higher than the primary, whereas peak dATP concentration was basically equivalent to the first Vpx + VLP treatment. Lastly, HIV-1 replication kinetics were faster in macrophages treated after the secondary Vpx treatments when compared to the initial single Vpx treatment.Conclusion: This study reveals that a very low level of SAMHD1 sufficiently modulates the normally low dNTP levels in macrophages and proposes potential diverse mechanisms of Vpx-mediated dNTP regulation in macrophages.",
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AU - Hollenbaugh, Joseph A.

AU - Tao, Sijia

AU - Lenzi, Gina M.

AU - Ryu, Sulryung

AU - Kim, Dong Hyun

AU - Diaz-Griffero, Felipe

AU - Schinazi, Raymond F.

AU - Kim, Baek

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AB - Background: SAMHD1 degrades deoxyribonucleotides (dNTPs), suppressing viral DNA synthesis in macrophages. Recently, viral protein X (Vpx) of HIV-2/SIVsm was shown to target SAMHD1 for proteosomal degradation and led to elevation of dNTP levels, which in turn accelerated proviral DNA synthesis of lentiviruses in macrophages.Results: We investigated both time-dependent and quantitative interplays between SAMHD1 level and dNTP concentrations during multiple exposures of Vpx in macrophages. The following were observed. First, SAMHD1 level was rapidly reduced by Vpx + VLP to undetectable levels by Western blot analysis. Recovery of SAMHD1 was very slow with less than 3% of the normal macrophage level detected at day 6 post Vpx treatment and only ~30% recovered at day 14. Second, dGTP, dCTP and dTTP levels peaked at day 1 post Vpx treatment, whereas dATP peaked at day 2. However, all dNTPs rapidly decreased starting at day 3, while SAMHD1 level was below the level of detection. Third, when Vpx pretreated macrophages were re-exposed to a second Vpx treatment at day 7, we observed dNTP elevation that had faster kinetics than the first Vpx + VLP treatment. Moreover, we performed a short kinetic analysis of the second Vpx treatment to find that dATP and dGTP levels peaked at 8 hours post secondary VLP treatment. dGTP peak was consistently higher than the primary, whereas peak dATP concentration was basically equivalent to the first Vpx + VLP treatment. Lastly, HIV-1 replication kinetics were faster in macrophages treated after the secondary Vpx treatments when compared to the initial single Vpx treatment.Conclusion: This study reveals that a very low level of SAMHD1 sufficiently modulates the normally low dNTP levels in macrophages and proposes potential diverse mechanisms of Vpx-mediated dNTP regulation in macrophages.

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