KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| RRM2 | Santa Cruz Biotechnology | Cat # sc-398294 |
| p16 | Abcam | Cat# ab108349; RRID:AB_10858268 |
| Vinculin | Sigma-Aldrich | Cat# V9131; RRID:AB_477629 |
| S6K | Cell Signaling Technology | Cat# 2708; RRID:AB_390722 |
| Phospho S6K (Thr389) | Cell Signaling Technology | Cat# 9234; RRID:AB_2269803 |
| 4E-BP1 | Cell Signaling Technology | Cat# 9644; RRID: AB_2097841 |
| Phospho 4E-BP1 (Ser65) | Cell Signaling Technology | Cat# 9451; RRID: AB_330947 |
| RPTOR | Cell Signaling Technology | Cat# 2280; RRID: AB_561245 |
| β-Actin | Sigma-Aldrich | Cat# A1978; RRID:AB_476692 |
| RPIA | Abcam | Cat# ab181235 |
| BRAF | Santa Cruz Biotechnology | Cat# sc-5284; RRID:AB_2721130 |
| RB | BD Biosciences | Cat# 554136; RRID:AB_39525 |
| CAD | Cell Signaling Technology | Cat# 11933; RRID: AB_2797772 |
| Phospho CAD (Ser1859) | Cell Signaling Technology | Cat # 70307; RRID: AB_2799782 |
| BrdU | Abcam | Cat# ab6326; RRID:AB_305426 |
| mTOR | Cell Signaling Technology | Cat# 2983; RRID:AB_2105622 |
| LAMP2 | Santa Cruz Biotechnology | Cat# sc-18822; RRID:AB_626858 |
| Anti-mouse HRP | Cell Signaling Technology | Cat# 7076; RRID:AB_330924 |
| Anti-rabbit HRP | Cell Signaling Technology | Cat# 7074; RRID:AB_2099233 |
| Anti-rat FITC | Jackson ImmunoResearch Labs | Cat# 712–095-150; RRID:AB_2340651 |
| Anti-rabbit FITC | Jackson ImmunoResearch Labs | Cat# 711–095-152; RRID:AB_2315776 |
| Anti-mouse Cy3 | Jackson ImmunoResearch Labs | Cat# 715–165-150; RRID:AB_2340813 |
| Bacterial and Virus Strains | ||
| Stbl3 Chemically Competent E. | Fisher Scientific | Cat# C737303 |
| DH5α Competent Cells | Fisher Scientific | Cat# 18265–017 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| BrdU | Alfa Aesar | Cat # H27260 |
| [35S]-methionine/cysteine | Perkin Elmer | Cat # NEG772 |
| X-Gal | Sigma-Aldrich | Cat # B4252 |
| D-Glucose-13C6 | Sigma-Aldrich | Cat # 389374 |
| Puromycin | GIBCO | Cat # A11138–02 |
| Polybrene | Sigma-Aldrich | Cat # H9268 |
| Propidium Iodide | Sigma-Aldrich | Cat# P4170 |
| PureProteome™ Protein G Magnetic Beads | EMD Millipore | Cat # LSKMAGG10 |
| cOmplete Mini EDTA-free | Roche | Cat # 11836170001 |
| Crystal violet | Harleco | Cat # 192–12 |
| DMEM 17 | Corning | Cat# 10–017-CV |
| DMEM 13 | Corning | Cat# 10–013-CV |
| RPMI | GIBCO | Cat# 11875093 |
| DMEM w/o glucose or glutamine | Sigma-Aldrich | Cat# D5030 |
| MEM/EBSS glutamine | HyClone | Cat# SH30024.01 |
| MEM Nonessential Amino Acids | Corning | Cat# 25025CL |
| Glutagro | Corning | Cat# 25015CL |
| Sodium Bicarbonate | Corning | Cat# 25035CL |
| Sodium Pyruvate | Corning | Cat# 25000CL |
| FBS | VWR | Cat# 16000–044 |
| Charcoal stripped FBS | Sigma-Aldrich | Cat# F6765 |
| Lipofectamine 2000 | Invitrogen | Cat# 11668019 |
| Trizol | Ambion | Cat# 15596018 |
| Glutaraldehyde | Polysciences, Inc. | Cat# 01909 |
| RNase Out | Invitrogen | Cat# 10777019 |
| Formaldehyde | VWR | Cat# 0493 |
| Paraformaldehyde | Sigma-Aldrich | Cat# 158127 |
| Temsirolimus | Selleckchem | Cat# S1044 |
| Torin 1 | Med Chem Express | Cat# HY-13003 |
| 7AAD | Tonbo | Cat# 13–6993-T500 |
| Optima LC-MS grade water | Thermo Fisher Scientific | Cat# W6–4 |
| Optima LC-MS grade acetonitrile | Thermo Fisher Scientific | Cat# A955–4 |
| Optima LC-MS grade methanol | Thermo Fisher Scientific | Cat# A456–4 |
| 1,1,1,3,3,3-hexafluoro 2-propanol (HFIP) | Sigma-Aldrich | Cat# 105228 |
| Diisopropylethylamine (DIPEA) | Sigma-Aldrich | Cat# D125806 |
| AMP standard | Sigma-Aldrich | Cat# 01930 |
| dAMP standard | Sigma-Aldrich | Cat# D6375 |
| ATP standard | Sigma-Aldrich | Cat# A26209 |
| dATP standard | Sigma-Aldrich | Cat# D6500 |
| dTMP standard | Sigma-Aldrich | Cat# T7004 |
| dTTP standard | Sigma-Aldrich | Cat# T0251 |
| CMP standard | Sigma-Aldrich | Cat# C1006 |
| dCMP standard | Sigma-Aldrich | Cat# D7625 |
| CTP standard | Sigma-Aldrich | Cat# C1506 |
| dCTP standard | Sigma-Aldrich | Cat# D4635 |
| UMP standard | Sigma-Aldrich | Cat# U6375 |
| dUMP standard | Sigma-Aldrich | Cat# D3876 |
| UDP standard | Sigma-Aldrich | Cat# 94330 |
| UTP standard | Sigma-Aldrich | Cat# U6625 |
| dUTP standard | Sigma-Aldrich | Cat# D4001 |
| AMP-13C,15N standard | Sigma-Aldrich | Cat# 650676 |
| dAMP-13C,15N standard | Sigma-Aldrich | Cat# 648620 |
| ATP-13C, 15N standard | Sigma-Aldrich | Cat# 645702 |
| dATP-13C, 15N standard | Sigma-Aldrich | Cat# 646237 |
| TTP-13C, 15N standard | Sigma-Aldrich | Cat# 646202 |
| CTP-13C, 15N standard | Sigma-Aldrich | Cat# 645699 |
| dTPC-13C, 15N standard | Sigma-Aldrich | Cat# 646229 |
| UTP-13C, 15N standard | Sigma-Aldrich | Cat# 645672 |
| dTMP-13C, 15N standard | Sigma-Aldrich | Cat# 648590 |
| CMP-13C, 15N standard | Sigma-Aldrich | Cat# 650692 |
| UMP-13C, 15N standard | Sigma-Aldrich | Cat# 651370 |
| Critical Commercial Assays | ||
| NucleoSpin RNA/Protein | Macherey-Nagel | Cat#740933 |
| Deposited Data | ||
| RNA-Seq | This paper | GEO: GSE133660 |
| Experimental Models: Cell Lines | ||
| Fibroblasts: IMR90 | ATCC | CCL-186 |
| Fibroblasts: IMR90 shControl | This paper | N/A |
| Fibroblasts: IMR90 shRRM2 | This paper | N/A |
| Fibroblasts: IMR90 shp16 | This paper | N/A |
| Fibroblasts: IMR90 shRRM2/shp16 | This paper | N/A |
| Fibroblasts: IMR90 shRRM2/shRB | This paper | N/A |
| Fibroblasts: IMR90 shRPIA | This paper | N/A |
| Fibroblasts: IMR90 shRRM2/shp16/shRPIA | This paper | N/A |
| Fibroblasts: IMR90 shRRM2/shp16/p16 OE | This paper | N/A |
| Fibroblasts: IMR90 Control | This paper | N/A |
| Fibroblasts: IMR90 BRAFV600E | This paper | N/A |
| Fibroblasts: IMR90 BRAFV600E/shp16 | This paper | N/A |
| Fibroblasts: IMR90 BRAFV600E/shRB | This paper | N/A |
| BRAFV600E/shp16/shRPIA | This paper | N/A |
| Embryonic kidney: 293FT | Thermo Fisher | R70007 |
| Embryonic kidney: Phoenix (QNX) | Dr. Gary Nolan | N/A |
| Skin: SKMel28 | ATCC | HTB-72 |
| Skin: SKMel28 shControl | This paper | N/A |
| Skin: SKMel28 shRPIA | This paper | N/A |
| Skin: SKMel28 shp16 | This paper | N/A |
| Skin: SKMel28 shp16/shRPIA | This paper | N/A |
| Skin: SKMel28 shRB | This paper | N/A |
| Skin: SKMel28: shp16/shRPTOR #1 | This paper | N/A |
| Skin: SKMel28: shp16/shRPTOR #2 | This paper | N/A |
| Pancreas: PATU8902 | G. DeNicola Laboratory | N/A |
| Pancreas: PATU8902 shControl | This paper | N/A |
| Pancreas: PATU8902 shRPIA | This paper | N/A |
| Pancreas: PATU8902 shp16 | This paper | N/A |
| Pancreas: PATU8902 shp16/shRPIA | This paper | N/A |
| Ovary: ES-2 | ATCC | Crl-1978 |
| Ovary: ES-2 shControl | This paper | N/A |
| Ovary: ES-2 shRPIA | This paper | N/A |
| Ovary: ES-2 shp16 | This paper | N/A |
| Ovary: ES-2 shp16/shRPIA | This paper | N/A |
| Bladder: TCC-SUP | ATCC | HTB-5 |
| Bladder: TCC-SUP shControl | This paper | N/A |
| Bladder: TCC-SUP shRPIA | This paper | N/A |
| Bladder: TCC-SUP shp16 | This paper | N/A |
| Bladder: TCC-SUP shp16/shRPIA | This paper | N/A |
| Colon: SW620 | ATCC | CCL-227 |
| Colon: SW620 shControl | This paper | N/A |
| Colon: SW620 shRPIA | This paper | N/A |
| Colon: SW620 shp16 | This paper | N/A |
| Colon: SW620 shp16/shRPIA | This paper | N/A |
| Colon: SW480 | ATCC | CCL-228 |
| Colon: SW480 shControl | This paper | N/A |
| Colon: SW480 shRPIA | This paper | N/A |
| Colon: SW480 shp16 | This paper | N/A |
| Colon: SW480 shp16/shRPIA | This paper | N/A |
| Colon: HT-29 | ATCC | HTB-38 |
| Colon: HT-29 shControl | This paper | N/A |
| Colon: HT-29 shRPIA | This paper | N/A |
| Colon: HT-29 shp16 | This paper | N/A |
| Colon: HT-29 shp16/shRPIA | This paper | N/A |
| Skin: SKMel28 shRB | This paper | N/A |
| Skin: SKMel28: shp16/shRPTOR #1 | This paper | N/A |
| Skin: SKMel28: shp16/shRPTOR #2 | This paper | N/A |
| Experimental Models: Organisms/Strains | ||
| C.B-17/ICRHsd-Prkdc scid mice | Envigo | 182 |
| Oligonucleotides | ||
| RT-qPCR Primers (Listed in Table S2) | This paper | N/A |
| Software and Algorithms | ||
| GSEA | Broad Institute | N/A |
| GraphPad Prism 7 | N/A | N/A |
| IDT tool for primer design | Integrated DNA Technologies | N/A |
| Cufflinks | Version v.2.0.2 | N/A |