Antibodies |
|
Anti-Strep-tag II antibody |
Abcam |
Cat#ab76949; RRID:AB_1524455
|
Anti-Histone H3 antibody |
Abcam |
Cat#ab10799; RRID:AB_470239
|
Anti-GFP from mouse IgG1κ (used for YFP detection) |
Sigma |
Cat#11814460001; RRID:AB_390913
|
GFP antibody rabbit polyclonal (used for YFP detection) |
Chromotek |
Cat#PABG1; RRID:AB_2749857
|
GAPDH (14C10) Rabbit mAb |
Cell Signaling |
Cat#2118; RRID:AB_561053
|
Anit-H1.10 antibody |
Abcam |
Cat#ab11079; RRID:AB_2295032
|
Anti-SPRTN mAB (6F2) |
Stingele lab |
Clone6F2 |
Goat Anti-Rat Immunoglobulins/HRP |
Sigma |
Cat#A9037; RRID:AB_258429
|
Goat Anti-Mouse Immunoglobulins/HRP |
Dako |
Cat#P0447; RRID:AB_2617137
|
Swine Anti-Rabbit Immunoglobulins/HRP |
Dako |
Cat#P0399; RRID:AB_2617141
|
Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 |
Thermo Scientific |
Cat#A-11001; RRID:AB_2534069
|
|
Bacterial and Virus Strains |
|
BL21(DE3) |
Thermo Scientific |
Cat#C600003 |
|
Chemicals, Peptides, and Recombinant Proteins |
|
16% Formaldehyde (w/v), Methanol-free |
Thermo Scientific |
Cat#28906 |
InstantBlue
|
Sigma |
Cat#ISB1L |
Doxycycline Hyclate |
Sigma |
Cat#D9891 |
ProLong Gold Antifade Reagent |
Thermo Fisher |
Cat#P10144
|
DAPI Solution |
Thermo Fisher |
Cat#62248 |
4x NuPAGE LDS sample buffer |
Thermo Scientific |
Cat#NP0007 |
Phusion HF enzyme |
NEB |
Cat#M0530 |
UltraPure BSA |
Thermo Scientific |
Cat#AM2616 |
Histone H1° Human |
NEB |
Cat#M2501S |
Protein G |
BioVision |
Cat#6510 |
Lipofectamine 2000 |
Thermo Scientific |
Cat#11668030 |
IGEPAL |
Sigma |
Cat#I8896 |
Biotin |
IBA Lifesciences |
Cat#2-1016-005 |
Pefabloc SC |
Merck |
Cat#11585916001 |
TCEP |
ROTH |
Cat#HN95.2 |
cOmplete EDTA-free protease inhibitor cocktail |
Merck |
Cat#4693132001 |
|
Critical Commercial Assays |
|
proFIRE Amine Coupling Kit |
Dynamic Biosensors |
Cat#PF-NH2-1 |
NucleoSpin® Gel and PCR Clean-up |
MACHEREY-NAGEL |
Cat#740609 |
Quant-iT PicoGreen dsDNA Assay Kit |
Thermo Scientific |
Cat#P11496
|
GeneJET Genomic DNA purification kit |
Thermo Scientific |
Cat#K0722 |
SsoAdvanced Universal SYBR Green Supermix |
Bio-Rad |
Cat#1725271 |
|
Experimental Models: Cell Lines |
|
Human HeLa Flp-In-T-REx |
The Francis Crick Institute Cell Services |
N/A |
Human HAP1 |
Thijn Brummelkamp, NKI Amsterdam |
N/A |
|
Oligonucleotides |
|
Oligonucleotide sequences used in this study are provided in Table S1
|
N/A |
N/A |
|
Recombinant DNA |
|
Ds ФX174 phage DNA RFI |
NEB |
Cat#N3021S |
Ss ФX174 phage DNA virion |
NEB |
Cat#N3023S |
pMAX-GFP |
LONZA |
Cat#VDC-1040 |
pNIC-STREP-ZB-SPRTN-WT |
This study |
N/A |
pNIC-STREP-ZB-SPRTN-Equation (E112Q) |
This study |
N/A |
pNIC-STREP-ZB-SPRTN-ZBD1∗ (Y179A_W197A) |
This study |
N/A |
pNIC-STREP-ZB-SPRTN-ZBD2∗ (R185A) |
This study |
N/A |
pNIC-STREP-ZB-SPRTN-BR∗ (K220A_K221E_G222A_K223A) |
This study |
N/A |
pNIC-STREP-ZB-SPRTN-EQ-ZBD1∗ (E112Q_Y179A_W197A) |
This study |
N/A |
pNIC-STREP-ZB-SPRTN-EQ-ZBD2∗ (E112Q_R185A) |
This study |
N/A |
pNIC-STREP-ZB-SPRTN-EQ-BR∗ (E112Q_ K220A_K221E_G222A_K223A) |
This study |
N/A |
pNIC-STREP-ZB-SPRTN-EQ-ZBD2∗-BR∗ (E112Q_R185A_K220A_K221E_G222A_K223A) |
This study |
N/A |
pNIC-STREP-ZB-SPRTN-ZBD-BR (aa151-245) |
This study |
N/A |
pNIC-STREP-ZB-SPRTN-ZBD (aa151-215) |
This study |
N/A |
pcDNA5-FRT/TO-YFP-SPRTN-WT-Strep |
Stingele et al., 2016 |
N/A |
pcDNA5-FRT/TO-YFP-SPRTN-Equation (E112Q)-Strep |
Stingele et al., 2016 |
N/A |
pcDNA5-FRT/TO-YFP-SPRTN-ZBD1∗ (Y179A_W197A)-Strep |
This study |
N/A |
pcDNA5-FRT/TO-YFP-SPRTN-ZBD2∗ (R185A)-Strep |
This study |
N/A |
pcDNA5-FRT/TO-YFP-SPRTN-BR∗ (K220A_K221E_G222A_K223A)-Strep |
This study |
N/A |
pOG44 |
Thermo Scientific |
Cat#V600520 |
pBABE-puro |
Addgene |
Cat#1764 |
pBABE-puro-SPRTN-WT-Strep |
This study |
N/A |
pBABE- puro-SPRTN-Equation (E112Q)-Strep |
This study |
N/A |
pBABE- puro-SPRTN-ZBD1∗ (Y179A_W197A)-Strep |
This study |
N/A |
pBABE- puro-SPRTN-ZBD2∗ (R185A)-Strep |
This study |
N/A |
|
Software and Algorithms |
|
Prism 7 |
GraphPad Software |
https://www.graphpad.com/ |
ImageJ |
NIH |
https://imagej.net/Fiji/Downloads |
Adobe Photoshop CC2018 |
Adobe |
https://www.adobe.com/es/products/photoshop.html |
|
Other |
|
HiTrap Heparin HP affinity columns |
GE Healthcare |
Cat#17040701 |
PD-10 Desalting columns |
GE Healthcare |
Cat#17085101 |
Strep-Tactin®XT Superflow® high capacity cartridges |
IBA Lifesciences |
Cat#2-4026-001 |
HiLoad 16/600 Superdex 200 pg column |
GE Healthcare |
Cat#GE28-9893-35 |
10 kDa cutoff Amicon Ultra centrifugal filters |
Merck |
Cat#UFC801096 |
GFP-Trap Magnetic Agarose |
Chromotek |
Cat#gtma-10 |