Table 1 Key reagents and resources
From: PIAS1 potentiates the anti-EBV activity of SAMHD1 through SUMOylation
Reagent or resource | Source | Identifier |
|---|---|---|
Antibodies and reagents | ||
 Anti-phospho-SAMHD1-T592 | Cell Signaling Tech | Cat# 15,038 |
 Anti-SAMHD1 | Bethyl | Cat# A311-354 |
 Anti-HA | Roche | Cat# 11–867-431–001 |
 Anti-HA-HRP | Cell Signaling Tech | Cat# 14,031 |
 SUMO2 Conjugation Kit | UBPBio | Cat# J3120 |
 SUMOlink SUMO2/3 Kit | Active motif | Cat# 40,220 (discontinued) |
 Anti-V5 magnetic beads | MBL | Cat# M167-11 |
 Anti-SUMO2/3 | MBL | Cat# M114-3 |
 Anti-SUMO2/3 | Active motif | Cat# 101,898 |
 Anti-V5-HRP | Thermo Fisher | Cat# R961-25 |
 Anit-V5 | Thermo Fisher | Cat# R960-25 |
 Anti-p53 | Active motif | Cat# 100,853 |
 Mouse anti-β-actin antibody | MP Biomedicals | Cat# 691,001 |
 Anti-human IgG (for IgG cross-linking) | MP Biomedicals | Cat# 55,087 |
 Anti-ZTA(BZ1) | Santa Cruz | Cat# sc-53904 |
 Rabbit IgG | Cell Signaling Tech | Cat# 2729 |
 Halo-tag protein purification kit | VWR/Promega | Cat# PAG6790 |
 Glutathione Sepharose 4B | GE Healthcare | Cat# 17–0756-01 |
 Factor Xa | NEB | Cat# P8010 |
 ChIP-Enzymatic Chromatin IP Kit | Cell Signaling Tech | Cat# 9003 |
Constructs | ||
 pLX304-SAMHD1 (with PAM mutated) | Li Lab [6] | pKZ175 |
 pLX304-SAMHD1-K469R/K595R/K622R (with PAM mutated) | This study | pSF029 |
 pLX304 | Addgene | Plasmid# 25,890 |
 LentiCRISPR v2 vector | Addgene | plasmid# 52,961 |
 pMD2.G | Addgene | plasmid# 12,259 |
 psPAX2 | Addgene | plasmid#12,260 |
 pSG5-ZTA | Hayward Lab Collection | NA |
 Halo-HA-BGLF4 | Hayward Lab Collection | pGL772 |
 Halo-HA-UL97 | Hayward Lab Collection | pGL798 |
 Halo-V5-PIAS1 | Li Lab [31] | pKZ28 |
 pCMV-XL4-SAMHD1 | Origene | Cat# SC114650 |
 pCMV-XL4-SAMHD1-K469R | This study | pSF004 |
 pCMV-XL4-SAMHD1-K595R | This study | pSF005 |
 pCMV-XL4-SAMHD1-K622R | This study | pSF006 |
 pGEX-5x-2-SAMHD1 | Li lab [6] | pKZ49 |
 pGEX-5x-2-SAMHD1-K469R | This study | pSF017 |
 pGEX-5x-2-SAMHD1-K595R | This study | pSF018 |
  pGEX-5x-2-SAMHD1-K622R | This study | pSF019 |
pGEX-5x-2-SAMHD1-K469R/K595R/K622R | This study | pSF028 |
Cell lines | ||
 Akata (EBV +) | Hayward Lab Collection | NA |
 Akata (EBV +)-PIAS1-KO | Li Lab [31] | NA |
 Akata (EBV +)-SAMHD1-sg1 | Li Lab [6] | NA |
 Akata (EBV +)-SAMHD1-sg1-pLX-SAMHD1 | This study | NA |
 Akata (EBV +)-SAMHD1-sg1-pLX-SAMHD1- K469R/K595R/K622R | This study | NA |
 Akata (EBV +)-SAMHD1-sg1-pLX-Vector | Li lab [6] | NA |
 293 T cells | Hayward Lab Collection | NA |
 HEK 293 (EBV +) | [50] | NA |