| Antibodies |
|
| GFP Polyclonal Antibody |
Thermo Fisher Scientific |
Cat# A-11122, RRID:AB_221569 |
| c-Myc Monoclonal Antibody (9E10) |
Thermo Fisher Scientific |
Cat# MA1-980, RRID:AB_558470 |
| Anti-mouse IgG, HRP-linked Antibody |
Cell Signaling Technology |
Cat# 7076, RRID:AB_330924 |
| Anti-rabbit IgG, HRP-linked Antibody |
Cell Signaling Technology |
Cat# 7074, RRID:AB_2099233 |
|
| Bacterial and virus strains |
|
|
E.coli BL21 (DE3) GOLD expression strain |
Agilent |
Cat #230132 |
|
| Chemicals, peptides, and recombinant proteins |
|
| ZnSO4
|
Fisher |
Z76-500 |
| PEG6K |
Hampton Research |
HR2-533 |
| β-D-thiogalactopyranoside (IPTG) |
Gold Bio |
Cat #367-93-1 |
| Complete Protease inhibitor tablets |
Roche |
Cat #14696200 |
| Kanamycin |
Gold Bio |
Cat #K-120-100 |
| Tris (2-carboxyethyl) phosphine hydrochloride (TCEP) |
Gold Bio |
Cat #TCEP-25 |
|
15N ammonium chloride 99% |
Cambridge Isotope Laboratories |
Cat #NLM-465-25 |
| HisTrap HP column |
GE Healthcare |
17524801 |
| Ni Sepharose 6 Fast flow 1000mL |
GE Healthcare |
Cat #17-5318-04 |
| HiLoad 26/60 Superdex 75 pg |
GE Healthcare |
Cat #17-1070-01 |
| Luria Broth |
Fisher |
BP9722-5 |
| Schneider’s Insect Medium |
Gibco |
Cat# 21720-024 |
| GFP-Trap magnetic beads |
Chromotek |
Cat# gtmak |
| Effectene transfection reagent |
Qiagen |
Cat# 301425 |
| Fetal Bovine Serum (Heat Inactivated) |
Gibco |
Cat# A5256801 |
| Penicillin-Streptomycin |
Sigma-Aldrich |
Cat# P0781 |
| RIPA buffer |
Thermo Fisher Scientific |
Cat# 89900 |
| SIGMAFAST Protease Inhibitor Tablets |
Sigma-Aldrich |
Cat# S8820 |
| Laemmli buffer |
Sigma-Aldrich |
Cat# S3401 |
| ECL Prime Western Blotting Detection Reagent |
Cytiva |
Cat# RPN2232 |
| DAPI |
Thermo Fisher Scientific |
Cat# 62248 |
| Hoechst 33342 |
Thermo Fisher Scientific |
Cat# 62249 |
| ProLong Diamond Antifade Mountant |
Thermo Fisher Scientific |
Cat# P36961
|
|
| Critical commercial assays |
|
| Stranded Total RNA Prep, Ligation with Ribo-Zero Plus |
Illumina |
Cat# 20040525 |
|
| Deposited data |
|
| PDB (Structure of PNUTS:Tox4 complex) |
This study |
PDBID 9CI7 |
| EMBL-EBI ArrayExpress (RNA sequencing data) |
This study |
E-MTAB-13735 |
|
| Oligonucleotides |
|
| See Table S5
|
|
|
|
| Recombinant DNA |
|
| pRP1b-(rat) PNUTS5–160
|
Peti and Page65
|
N/A |
| pTHMT-(human) Tox4571–621
|
This study |
N/A |
| pAGW-Drosophila tox4wt
|
This study |
N/A |
| pAGW-Drosophila tox4P216Ter
|
This study |
N/A |
| pAW-Drosophila PNUTSwt
|
This study |
N/A |
| pCaSpeR4-attB, GFP-Drosophila tox4wt
|
This study |
N/A |
| pCaSpeR4-attB, GFP-Drosophila tox4P216Ter
|
This study |
N/A |
| pw8-attB, Drosophila PNUTSwt-GFP |
This study |
N/A |
| pw8-attB, Drosophila PNUTSK42E,V44D-GFP |
This study |
N/A |
| pw8-attB, Drosophila PNUTSL10E,K42E,V44D-GFP |
This study |
N/A |
| pw8-attB, Drosophila pNUTSwt-flp-W726A
|
This study |
N/A |
|
| Software and algorithms |
|
| Topspin 3.5/4.0.5 |
Bruker |
RRID:SCR_014227 |
| solve_structure script |
SSRL |
http://smb.slac.stanford.edu/facilities/software/MAD_scripts
|
| XDS |
Kabsch et al.66
|
https://www.ccp4.ac.uk/
|
| Truncate |
French and Wilson67
|
https://www.ccp4.ac.uk/
|
| Aimless |
Evans and Murshudov68
|
RRID:SCR_015747 |
| Solve/Resolve |
Terwilliger and Berendzen 69
|
https://solve.lanl.gov/
|
| PHENIX |
Zwart et al.70
|
RRID:SCR_014224 |
| COOT |
Emsley et al.71
|
RRID:SCR_014222 |
| Pymol |
Schrodinger, LLC |
RRID:SCR_000305 |
| NITPIC |
Scheuermann et al.72
|
https://www.utsouthwestern.edu/labs/mbr/software/
|
| SEDPHAT |
Zhao et al.73
|
https://www.utsouthwestern.edu/labs/mbr/software/
|
| GUSSI |
Scheuermann et al.72
|
https://www.utsouthwestern.edu/labs/mbr/software/
|
| FIJI (ImageJ) |
NIH |
RRID:SCR_002285 |
| Zen (v 3.9) |
Zeiss |
RRID:SCR_013672 |
| Imaris (v 10) |
Bitplane/Oxford Instruments |
RRID:SCR_007370 |
| Prism (v 10) |
Graphpad |
RRID:SCR_002798 |
| FastQC |
Babraham Bioinformatics |
RRID:SCR_014583 |
| DESeq2 |
Love et al.74
|
RRID:SCR_015687 |
| Trimmomatic |
Bolger et al.75
|
RRID:SCR_011848 |
| STAR |
Dobin et al.76
|
RRID:SCR_004463 |
| Metascape |
Zhou et al.77
|
RRID:SCR_016620 |
| FlyEnrichR |
Kuleschov et al.47
|
https://maayanlab.doud/FlyEnrichr
|
|
| Other |
|
| Cell Discoverer 7 with 20× objective |
Zeiss |
N/A |
| LSM880 with 20× objective |
Zeiss |
N/A |
| A1R confocal with 20× Plan-Apo objective |
Nikon |
N/A |
| 24 well Sensoplate |
Greiner |
Cat# 662892 |
| Coverslips No 1 |
Scientific Laboratory Supplies |
Cat# MIC3110 |
| ChemiDoc MP Imaging System |
Biorad |
12003154 |
