KEY RESOURCES TABLE

REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Anti-PNPT1 antibody	Proteintech	Cat# 14487-1-AP; RRID: AB_2165820
Anti-PNPT1 monoclonal antibody	Invitrogen	Cat# MA5-27859; RRID: AB_2735325
Anti-PNPT1 antibody	Abcam	Cat# ab157109; RRID: AB_2910229
Anti-MGMT antibody	Proteintech	Cat# 17195-1-AP; RRID: AB_2143221
Anti-MGMT monoclonal antibody	Invitrogen	Cat# 35-7000; RRID: AB_87574
Anti-MGMT antibody	Abcam	Cat# ab39253; RRID: AB_776338
Anti-Alpha (α) tubulin antibody	Santa Cruz	Cat# sc-8035; RRID: AB_628408
Anti-GFP monoclonal antibody	Santa Cruz	Cat# sc-9996; RRID: AB_627695
Anti-GFP antibody	Proteintech	Cat# 50430-2-AP; RRID: AB_11042881
Anti-GFP antibody	OriGene	Cat# TA150039; RRID: AB_2622255
Anti-GAPDH antibody	Proteintech	Cat# 10494-1-AP; RRID: AB_2263076
Anti-Succinate dehydrogenase antibody	Proteintech	Cat# 14865-1-AP; RRID: AB_11182164
Anti-FLAG monoclonal antibody	Millipore-Sigma	Cat# F3165; RRID: AB_259529
Anti-IgG mouse	Cell Signaling	Cat# 7074; RRID: AB_2099233
Anti-IgG mouse	Santa Cruz	Cat# sc-2025; RRID: AB_737182
Anti-IgG rabbit	Cell Signaling	Cat# 7076; RRID: AB_330924
Bacterial and virus strains
DH5α	Invitrogen	18265017
XL1-Blue	Agilent	200150
Stbl3	Invitrogen	C737303
Chemicals, peptides, and recombinant proteins
Temozolomide	Sigma-Aldrich	T2577
Actinomycin D1	Gibco	11805017
VE821	MCE	HY-14731
TRIzol reagent	Invitrogen	15596026
DMSO	Sigma-Aldrich	D2438
Dynabeads M-280 Streptavidin	Invitrogen	11206D
Dynabeads Protein G	Invitrogen	10009D
Dynabeads Protein A	Invitrogen	10008D
Ketamine	Dechra	200–073
Xylazine	Dechra	047–956
Heparin Solution	STEMCELL Technologies	07980
Human Epidermal Growth Factor	STEMCELL Technologies	78006
Human Fibroblast Growth Factor	STEMCELL Technologies	78134
Puromycin	Sigma-Aldrich	P9620
Neomycin	Sigma-Aldrich	N1142
Doxycycline	Sigma-Aldrich	D3447
D-Luciferin	Thermo Fisher	L2916
Critical chemical assays
Dual-Glo® Luciferase Assay System	Promega	E2920
miRNeasy Kit	Qiagen	217084
OmniScript RT Kit	Qiagen	205113
miRCURY LNA RT Kit	Qiagen	339340
QuikChange Lightning Site-Directed mutagenesis Kit	Agilent	210519
Bioanalyzer RNA 6000 Nano Kit	Agilent	5067–1511
Rabbit EnVision™ Kit	Dako	K4003
DNeasy blood and tissue Kit	Qiagen	69504
EpiTect bisulfite Kit	Qiagen	59104
Mitochondria isolation Kit	Thermo Fisher	89874
Universal Mycoplasma Detection Kit	ATCC	30-1012K
Deposited data
RNA Sequencing of BT-83 and CMK3 cell lines	This paper	DOI: https://doi.org/10.17632/pdvttwws8m.1
Experimental models: Cell lines
CMK3	This paper	N/A
BT-83	This paper	N/A
Experimental models: Organisms/strains
Wild-type nude mice	The Jackson Laboratory	Strain #002019
Oligonucleotides
PNPT1 For: 5′TGTCATGTTGGAAGCCTCTG3′	IDT	N/A
PNPT1 Rev: 5′ACTGCTGAATGCCCTGAATTA3′	IDT	N/A
PNPT1-Myc For: 5′CAGTCGCCAGCTACAACCGTGGT3′	IDT	N/A
PNPT1-Myc Rev: 5′CTCTTCTGAGATGAGTTTCTGCTC3′	IDT	N/A
MGMT For: 5′GCCTGGCTGAATGCCTATTT3′	IDT	N/A
MGMT Rev: 5′AACCTTCAGCTTCCATAAC3′	IDT	N/A
PNPT1-MRE2: 5′ CGCGTAGCCCAGAAGT TCGACACTGTAGTGGC3′	IDT	N/A
PNPT1-MRE2 Rev: 5′ GGCCGCGACTACAGT GTCGAACTTCTGGGCTA3′	IDT	N/A
PNPT1-MRE3: 5′ CGCGTGACCAGCCTGGC CAACATGGTGAAAGC3′	IDT	N/A
PNPT1-MRE3 Rev: 5′ GGCCGCTTTCACCA TGTTCGCCAGGCTGGTCA3′	IDT	N/A
PNPT1-MRE2-mut For: 5′ CGCGTAGAAAA GAAGTTCGACACTGTAGTGGC3′	IDT	N/A
PNPT1-MRE2-mut Rev: 5′ GGCCGCGACT ACAGTGTCGAACTTCTTTTCTA3′	IDT	N/A
PNPT1-MRE3-mut For: 5′ CGCGTGACCA TAATGGCCAACATGGTGAAAGC3′	IDT	N/A
PNPT1-MRE3-mut Rev: 5′ GGCCGCTTTC ACCATGTTCGCCATTATGGTCA3′	IDT	N/A
PNPT1 S135G For: 5′ATTGGTACTGGTGATAAAGAA3′	IDT	N/A
PNPT1 S135G Rev: 5′TTCTTTATCACCAGTACCAAT3′	IDT	N/A
PNPT1 Q387R For: 5′GCATTATTTCGAAGAGGACAA3′	IDT	N/A
PNPT1 Q387R Rev: 5′TTGTCCTCTTCGAAATAATGC3′	IDT	N/A
PNPT1 R445E For: 5′GGTTTAAATGAAAGAGAACTT3′	IDT	N/A
PNPT1 R445E Rev: 5′AAGTTCTCTTTCATTTAAACC3′	IDT	N/A
PNPT1 R446E For: 5′TTAAATAGAGAAGAACTTGGG3′	IDT	N/A
PNPT1 R446E Rev: 5′CCCAAGTTCTTCTCTATTTAA3′	IDT	N/A
PNPT1 S484A For: 5′GGGTCATCTGCTATGGCATCT3′	IDT	N/A
PNPT1 S484A Rev: 5′AGATGCCATAGCAGATGACCC3′	IDT	N/A
PNPT1 D538A For: 5′GGAATTGAAGCTTACAATGGT3′	IDT	N/A
PNPT1 D538A Rev: 5′ACCATTGTAAGCTTCAATTCC3′	IDT	N/A
PNPT1 D544G For: 5′GGTGACATGGGCTTCAAAATA3′	IDT	N/A
PNPT1 D544G Rev: 5′TATTTTGAAGCCCATGTCACC3′	IDT	N/A
Unmet MGMT For: 5′TTTGTGTTTTGATGTTTGTAGGTTTTTGT3′	IDT	N/A
Unmet MGMT Rev: 5′AACTCCACACTCT TCCAAAAACAAAACA3′	IDT	N/A
Met MGMT For: 5′TTTCGACGTTCGTAG GTTTTCGC3′	IDT	N/A
Met MGMT Rev: 5′GCACTCTTCCGAAA ACGAAACG3′	IDT	N/A
Recombinant DNA
pLenti-CRISPR v2-sgPNPT1-2	Addgene	234770
pCMV-Entry-miR-181d	This paper	N/A
pCMV-VSV-G	Addgene	8454
psPAX2	Addgene	12260
pCMV6-Entry-PNPT1	OriGene	RC208800
pCMV6-AC-GFP-PNPT1	OriGene	RG208800
pCMV6-AC-GFP-PNPT1 S135G	This paper	N/A
pCMV6-AC-GFP-PNPT1 Q387R	This paper	N/A
pCMV6-AC-GFP-PNPT1 R445E	This paper	N/A
pCMV6-AC-GFP-PNPT1 R446E	This paper	N/A
pCMV6-AC-GFP-PNPT1 S484A	This paper	N/A
pCMV6-AC-GFP-PNPT1 D538A	This paper	N/A
pCMV6-AC-GFP-PNPT1 D544G	This paper	N/A
psiCHECK-2	Promega	N/A
psiCHECK-2-PNPT1 MRE1	This paper	N/A
psiCHECK-2-PNPT1 MRE2	This paper	N/A
psiCHECK-2-PNPT1 MRE3	This paper	N/A
psiCHECK-2-PNPT1 mut-MRE2	This paper	N/A
psiCHECK-2-PNPT1 mut-MRE3	This paper	N/A
pCMV6-Entry-PNPT1-MRE2	This paper	N/A
pCMV6-Entry-PNPT1-MRE3	This paper	N/A
pCMV6-Entry-PNPT1-MRE2-mut	This paper	N/A
pCMV6-Entry-PNPT1-MRE3-mut	This paper	N/A
pINDUCER20_3xFlag	Addgene	190028
pINDUCER20_3xFlag-shATR	This paper	N/A
pLV-fLuc/puro	This paper	Gifted by Dr. Robert Galvin
Software and algorithms
Graphpad Prsim	Graphpad	Version 10.0
ImageJ	NIH	Version 1.50e
Adobe photoshop	Adobe	Version 21.0.2