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Table 1 SAMHD1 oligomerization variants

From: Contribution of oligomerization to the anti-HIV-1 properties of SAMHD1

SAMHD1 variant

HIV-1 Restrictiona

Oligomerizationb

RNA Bindingc

Localizationd

Cellular dNTP Levele

Association with WT ± SDf

Self-association ± SDg

WT

+

+

+

N

Low

100

100

Y146S/Y154S

+

-

+

N

Low

10.15 ± 1.48

2.46 ± 0.69

L428S/Y432S

+

-

+

N

Low

25.9 ± 9.19

16.04 ± 1.50

Y146S/Y154S/L428S/Y432S

ND

-

+

N/C

Low

1.4 ± 0.25

22.04 ± 7.70

  1. aHIV-1 restriction was measured by infecting U937 cells stably expressing the indicated SAMHD1 variants with HIV-1-GFP. After 48 hours, the percentage of GFP-positive cells (infected cells) was determined by flow cytometry.
  2. bOligomerization of the different SAMHD1 variants was determined by measuring the ability of the SAMHD1-FLAG variant to interact with wild type SAMHD1-HA variant, as described [31]. “ + ” indicates 100% oligomerization, which corresponds to the amount of wild type SAMHD1-HA that interacts with wild type SAMHD1-FLAG. “ - ” indicates the absence of oligomerization.
  3. cSAMHD1-FLAG variants were assayed for their ability to bind the double-stranded RNA analog ISD-PS, as described [31]. “+” indicates the RNA binding achieved by wild type SAMHD1.
  4. dSubcellular localization of the different SAMHD1 variants in HeLa cells was performed as described [31]. “N” indicates nuclear localization; “N/C” indicates nuclear and cytoplasmic localization.
  5. eThe cellular dATP levels of PMA-treated U937 cells stably expressing the different SAMHD1 variants were determined by primer extension as described [31]. “Low” indicates similar to the dATP levels observed in PMA-treated U937 cells stably expressing wild type SAMHD1.
  6. fWT and SAMHD1-FLAG variants were assayed for association with wild-type SAMHD1-HA as described [31]. Percentages are an average of two independent experiments. The percentage represents the fraction of the SAMHD1 variant coprecipitated with wild-type SAMHD1 relative to the amount of wild-type SAMHD1 coprecipitated with itself.
  7. gWT and SAMHD1-FLAG variants were assayed for association with wild-type and variant SAMHD1-HA as described [33]. Percentages are an average of two independent experiments. The percentage represents the fraction of the SAMHD1 variant coprecipitated with itself relative to the coprecipitation of wild-type SAMHD1 with itself.
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Retrovirology

ISSN: 1742-4690

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