AMPK regulation of Raptor and TSC2 mediate metformin effects on transcriptional control of anabolism and inflammation.
Jeanine L Van Nostrand,
Kristina Hellberg,
En-Ching Luo,
Eric L Van Nostrand,
Alina Dayn,
Jingting Yu,
Maxim N Shokhirev,
Yelena Dayn,
Gene W Yeo,
Reuben J Shaw
Genes & development
· 2020
· Vol. 34
(19-20)
· pp. 1330-1344
Abstract
Despite being the frontline therapy for type 2 diabetes, the mechanisms of action of the biguanide drug metformin are still being discovered. In particular, the detailed molecular interplays between the AMPK and the mTORC1 pathway in the hepatic benefits of metformin are still ill defined. Metformin-dependent activation of AMPK classically inhibits mTORC1 via TSC/RHEB, but several lines of evidence suggest additional mechanisms at play in metformin inhibition of mTORC1. Here we investigated the role of direct AMPK-mediated serine phosphorylation of RAPTOR in a new <i>Raptor</i><sup><i>AA</i></sup> mouse model, in which AMPK phospho-serine sites Ser722 and Ser792 of RAPTOR were mutated to alanine. Metformin treatment of primary hepatocytes and intact murine liver requires AMPK regulation of both RAPTOR and TSC2 to fully inhibit mTORC1, and this regulation is critical for both the translational and transcriptional response to metformin. Transcriptionally, AMPK and mTORC1 were both important for regulation of anabolic metabolism and inflammatory programs triggered by metformin treatment. The hepatic transcriptional response in mice on high-fat diet treated with metformin was largely ablated by AMPK deficiency under the conditions examined, indicating the essential role of this kinase and its targets in metformin action in vivo.
Publication Types
["Journal Article", "Research Support, N.I.H., Extramural", "Research Support, Non-U.S. Gov't"]
MeSH Terms
["AMP-Activated Protein Kinases", "Animals", "Diabetes Mellitus, Type 2", "Disease Models, Animal", "Gene Expression Regulation", "Gene Knock-In Techniques", "Genotype", "Hypoglycemic Agents", "Inflammation", "Mechanistic Target of Rapamycin Complex 1", "Metabolism", "Metformin", "Mice", "Phosphorylation", "Regulatory-Associated Protein of mTOR", "Signal Transduction", "TOR Serine-Threonine Kinases", "Tuberous Sclerosis Complex 2 Protein"]
Funding
P30 CA014195
NCI NIH HHS
(United States)
K99 HG009530
NHGRI NIH HHS
(United States)
R01 DK080425
NIDDK NIH HHS
(United States)
P01 CA120964
NCI NIH HHS
(United States)
R35 CA220538
NCI NIH HHS
(United States)
Linked Datasets (4)
RNA-seq of metformin treatment in primary murine hepatocytes in WT, Raptor Ser-Ala mutant, Tsc2-null, Raptor mutant;Tsc2-null, and Ampk-null cells
Mus musculus65 data files
| File | Type | Size |
|---|---|---|
| AAIns_JVN10_S10_R1_001.fastq.gz | RNA-Seq | 320.1 MB |
| AAIns_JVN11_S11_R1_001.fastq.gz | RNA-Seq | 332.6 MB |
| AAIns_JVN9_S9_R1_001.fastq.gz | RNA-Seq | 315.9 MB |
| AAInsMet_JVN12_S12_R1_001.fastq.gz | RNA-Seq | 296.3 MB |
| AAInsMet_JVN13_S13_R1_001.fastq.gz | RNA-Seq | 288.6 MB |
| AAInsMet_JVN14_S14_R1_001.fastq.gz | RNA-Seq | 378.9 MB |
| AAMet_JVN16_S16_R1_001.fastq.gz | RNA-Seq | 305.6 MB |
| AAMet_JVN7_S61_R1_001.fastq.gz | RNA-Seq | 361.6 MB |
| AAMet_JVN8_S62_R1_001.fastq.gz | RNA-Seq | 413.5 MB |
| AATSCIns_JVN25_S25_R1_001.fastq.gz | RNA-Seq | 388.2 MB |
| AATSCIns_JVN26_S26_R1_001.fastq.gz | RNA-Seq | 370.2 MB |
| AATSCIns_JVN27_S27_R1_001.fastq.gz | RNA-Seq | 358.1 MB |
| AATSCInsMET_JVN28_S28_R1_001.fastq.gz | RNA-Seq | 379.4 MB |
| AATSCInsMET_JVN29_S29_R1_001.fastq.gz | RNA-Seq | 375.3 MB |
| AATSCInsMET_JVN30_S30_R1_001.fastq.gz | RNA-Seq | 384.6 MB |
| AATscMet_JVN15_S69_R1_001.fastq.gz | RNA-Seq | 398.3 MB |
| AATscMet_JVN16_S70_R1_001.fastq.gz | RNA-Seq | 392.2 MB |
| AATscMet_JVN32_S32_R1_001.fastq.gz | RNA-Seq | 368.6 MB |
| AATscUT_JVN13_S67_R1_001.fastq.gz | RNA-Seq | 348.3 MB |
| AATscUT_JVN14_S68_R1_001.fastq.gz | RNA-Seq | 385.0 MB |
| AATscUT_JVN31_S31_R1_001.fastq.gz | RNA-Seq | 413.6 MB |
| AAUT_JVN15_S15_R1_001.fastq.gz | RNA-Seq | 318.2 MB |
| AAUT_JVN5_S59_R1_001.fastq.gz | RNA-Seq | 341.8 MB |
| AAUT_JVN6_S60_R1_001.fastq.gz | RNA-Seq | 353.5 MB |
| AMPKIns_JVN33_S33_R1_001.fastq.gz | RNA-Seq | 434.1 MB |
| AMPKIns_JVN34_S34_R1_001.fastq.gz | RNA-Seq | 388.5 MB |
| AMPKIns_JVN35_S35_R1_001.fastq.gz | RNA-Seq | 400.4 MB |
| AMPKInsMet_JVN36_S36_R1_001.fastq.gz | RNA-Seq | 366.2 MB |
| AMPKInsMet_JVN37_S37_R1_001.fastq.gz | RNA-Seq | 372.7 MB |
| AMPKInsMet_JVN38_S38_R1_001.fastq.gz | RNA-Seq | 336.8 MB |
| AmpkMet_JVN19_S73_R1_001.fastq.gz | RNA-Seq | 345.8 MB |
| AmpkMet_JVN20_S74_R1_001.fastq.gz | RNA-Seq | 328.8 MB |
| AMPKMet_JVN40_S40_R1_001.fastq.gz | RNA-Seq | 357.5 MB |
| AmpkUT_JVN17_S71_R1_001.fastq.gz | RNA-Seq | 359.1 MB |
| AmpkUT_JVN18_S72_R1_001.fastq.gz | RNA-Seq | 380.4 MB |
| AmpKUT_JVN39_S39_R1_001.fastq.gz | RNA-Seq | 356.0 MB |
| TscIns_JVN17_S17_R1_001.fastq.gz | RNA-Seq | 357.6 MB |
| TscIns_JVN18_S18_R1_001.fastq.gz | RNA-Seq | 340.7 MB |
| TscIns_JVN19_S19_R1_001.fastq.gz | RNA-Seq | 355.8 MB |
| TscInsMet_JVN20_S20_R1_001.fastq.gz | RNA-Seq | 349.4 MB |
| TscInsMet_JVN21_S21_R1_001.fastq.gz | RNA-Seq | 335.5 MB |
| TscInsMet_JVN22_S22_R1_001.fastq.gz | RNA-Seq | 364.1 MB |
| TscMet_JVN11_S65_R1_001.fastq.gz | RNA-Seq | 329.4 MB |
| TscMet_JVN12_S66_R1_001.fastq.gz | RNA-Seq | 338.5 MB |
| TscMet_JVN24_S24_R1_001.fastq.gz | RNA-Seq | 328.1 MB |
| TscUT_JVN10_S64_R1_001.fastq.gz | RNA-Seq | 349.4 MB |
| TscUT_JVN23_S23_R1_001.fastq.gz | RNA-Seq | 376.3 MB |
| TscUT_JVN9_S63_R1_001.fastq.gz | RNA-Seq | 352.3 MB |
| WTInk_JVN21_S75_R1_001.fastq.gz | RNA-Seq | 307.2 MB |
| WTInk_JVN22_S76_R1_001.fastq.gz | RNA-Seq | 345.8 MB |
| WTInk_JVN41_S41_R1_001.fastq.gz | RNA-Seq | 349.5 MB |
| WTInsInk_JVN43_S43_R1_001.fastq.gz | RNA-Seq | 390.2 MB |
| WTIns_JVN1_S1_R1_001.fastq.gz | RNA-Seq | 296.2 MB |
| WTIns_JVN2_S2_R1_001.fastq.gz | RNA-Seq | 327.4 MB |
| WTIns_JVN3_S3_R1_001.fastq.gz | RNA-Seq | 324.9 MB |
| WTIns_JVN42_S42_R1_001.fastq.gz | RNA-Seq | 404.7 MB |
| WTInsMet_JVN4_S4_R1_001.fastq.gz | RNA-Seq | 317.0 MB |
| WTInsMet_JVN5_S5_R1_001.fastq.gz | RNA-Seq | 311.2 MB |
| WTInsMet_JVN6_S6_R1_001.fastq.gz | RNA-Seq | 329.2 MB |
| WTMet_JVN3_S57_R1_001.fastq.gz | RNA-Seq | 358.2 MB |
| WTMet_JVN4_S58_R1_001.fastq.gz | RNA-Seq | 362.8 MB |
| WTMet_JVN8_S8_R1_001.fastq.gz | RNA-Seq | 326.1 MB |
| WTUT_JVN1_S55_R1_001.fastq.gz | RNA-Seq | 343.4 MB |
| WTUT_JVN2_S56_R1_001.fastq.gz | RNA-Seq | 323.1 MB |
| WTUT_JVN7_S7_R1_001.fastq.gz | RNA-Seq | 346.9 MB |
RNA-seq of metformin treatment in liver in WT, Raptor Ser-Ala mutant, Tsc2-null, Raptor mutant;Tsc2-null, and Ampk-null
Mus musculus29 data files
| File | Type | Size |
|---|---|---|
| 1038_TSCSal_JVN13_S13_R1_001.fastq.gz | RNA-Seq | 790.1 MB |
| 1039_ATSal_JVN18_S18_R1_001.fastq.gz | RNA-Seq | 838.9 MB |
| 1069_ATSal_JVN19_S19_R1_001.fastq.gz | RNA-Seq | 728.5 MB |
| 1071_ATMet_JVN23_S23_R1_001.fastq.gz | RNA-Seq | 979.7 MB |
| 1087_WTMet_JVN6_S6_R1_001.fastq.gz | RNA-Seq | 1.1 GB |
| 1202_AAMet_JVN12_S12_R1_001.fastq.gz | RNA-Seq | 967.4 MB |
| 1268_AASal_JVN8_S8_R1_001.fastq.gz | RNA-Seq | 931.8 MB |
| 1269_AASal_JVN9_S9_R1_001.fastq.gz | RNA-Seq | 1004.6 MB |
| 1277_AAMet_JVN10_S10_R1_001.fastq.gz | RNA-Seq | 1.0 GB |
| 1289_AASal_JVN7_S7_R1_001.fastq.gz | RNA-Seq | 894.8 MB |
| 1291_TSCMet_JVN17_S17_R1_001.fastq.gz | RNA-Seq | 857.9 MB |
| 1362_WTSal_JVN3_S3_R1_001.fastq.gz | RNA-Seq | 835.9 MB |
| 1364_TSCSal_JVN14_S14_R1_001.fastq.gz | RNA-Seq | 859.1 MB |
| 1367_AAMet_JVN11_S11_R1_001.fastq.gz | RNA-Seq | 926.2 MB |
| 1369_TSCMet_JVN15_S15_R1_001.fastq.gz | RNA-Seq | 891.1 MB |
| 1373_TSCMet_JVN16_S16_R1_001.fastq.gz | RNA-Seq | 800.5 MB |
| 1406_ATSal_JVN20_S20_R1_001.fastq.gz | RNA-Seq | 850.5 MB |
| 1407_ATMet_JVN21_S21_R1_001.fastq.gz | RNA-Seq | 1.0 GB |
| 1408_ATMet_JVN22_S22_R1_001.fastq.gz | RNA-Seq | 1.0 GB |
| 4330_AdelMet_JVN29_S29_R1_001.fastq.gz | RNA-Seq | 809.5 MB |
| 4354_WTSal_JVN1_S1_R1_001.fastq.gz | RNA-Seq | 697.5 MB |
| 4355_AdelSal_JVN24_S24_R1_001.fastq.gz | RNA-Seq | 958.5 MB |
| 4356_AdelSal_JVN25_S25_R1_001.fastq.gz | RNA-Seq | 1.0 GB |
| 4357_WTSal_JVN2_S2_R1_001.fastq.gz | RNA-Seq | 877.5 MB |
| 4359_WTMet_JVN4_S4_R1_001.fastq.gz | RNA-Seq | 987.2 MB |
| 4360_WTMet_JVN5_S5_R1_001.fastq.gz | RNA-Seq | 976.5 MB |
| 4363_AdelSal_JVN26_S26_R1_001.fastq.gz | RNA-Seq | 997.9 MB |
| 4364_AdelMet_JVN27_S27_R1_001.fastq.gz | RNA-Seq | 908.1 MB |
| 4365_AdelMet_JVN28_S28_R1_001.fastq.gz | RNA-Seq | 819.0 MB |
Ribosome profiling of metformin treatment in primary murine hepatocytes in WT, Raptor Ser-Ala mutant, Tsc2-null, Raptor mutant;Tsc2-null, and Ampk-null cells
Mus musculus88 data files
| File | Type | Size |
|---|---|---|
| AAInsInput_JVN15_S31_R1_001.fastq.gz | RNA-Seq | 361.5 MB |
| aa_ins_input_JVN_64_S14_R1_001.fastq-2.gz | RNA-Seq | 373.2 MB |
| aa_ins_input_JVN_64_S14_R1_001.fastq.gz | RNA-Seq | 130.0 MB |
| AAIns_JVN31_S47_R1_001.fastq.gz | OTHER | 408.8 MB |
| aa_ins_JVN_53_S3_R1_001.fastq-2.gz | OTHER | 345.9 MB |
| aa_ins_JVN_53_S3_R1_001.fastq.gz | OTHER | 148.5 MB |
| AAInsMetInput_JVN16_S32_R1_001.fastq.gz | RNA-Seq | 349.8 MB |
| aa_ins_met_input_JVN_65_S15_R1_001.fastq-2.gz | RNA-Seq | 327.3 MB |
| aa_ins_met_input_JVN_65_S15_R1_001.fastq.gz | RNA-Seq | 111.9 MB |
| AAInsMet_JVN32_S48_R1_001.fastq.gz | OTHER | 429.8 MB |
| aa_ins_met_JVN_54_S4_R1_001.fastq-2.gz | OTHER | 330.7 MB |
| aa_ins_met_JVN_54_S4_R1_001.fastq.gz | OTHER | 143.2 MB |
| AAMetinput_JVN5_S21_R1_001.fastq.gz | RNA-Seq | 334.7 MB |
| AAMet_JVN30_S46_R1_001.fastq.gz | OTHER | 450.6 MB |
| AATSCInsInput_JVN19_S35_R1_001.fastq.gz | RNA-Seq | 393.6 MB |
| aa_tsc_ins_input_JVN_68_S18_R1_001.fastq-2.gz | RNA-Seq | 356.6 MB |
| aa_tsc_ins_input_JVN_68_S18_R1_001.fastq.gz | RNA-Seq | 121.9 MB |
| AATSCIns_JVN39_S55_R1_001.fastq.gz | OTHER | 391.6 MB |
| aa_tsc_ins_JVN_57_S7_R1_001.fastq-2.gz | OTHER | 275.0 MB |
| aa_tsc_ins_JVN_57_S7_R1_001.fastq.gz | OTHER | 118.9 MB |
| AATscInsMetInput_JVN20_S36_R1_001.fastq.gz | RNA-Seq | 304.4 MB |
| aa_tsc_ins_met_input_JVN_69_S19_R1_001.fastq-2.gz | RNA-Seq | 330.5 MB |
| aa_tsc_ins_met_input_JVN_69_S19_R1_001.fastq.gz | RNA-Seq | 113.5 MB |
| AATscInsMet_JVN40_S56_R1_001.fastq.gz | OTHER | 372.1 MB |
| aa_tsc_ins__met_JVN_58_S8_R1_001.fastq-2.gz | OTHER | 370.9 MB |
| aa_tsc_ins__met_JVN_58_S8_R1_001.fastq.gz | OTHER | 166.8 MB |
| AATscMetInput_JVN9_S25_R1_001.fastq.gz | RNA-Seq | 350.9 MB |
| AATscMet_JVN38_S54_R1_001.fastq.gz | OTHER | 416.0 MB |
| AATscUTInput_JVN8_S24_R1_001.fastq.gz | RNA-Seq | 349.6 MB |
| AATscUT_JVN37_S53_R1_001.fastq.gz | OTHER | 399.9 MB |
| AAUTinput_JVN4_S20_R1_001.fastq.gz | RNA-Seq | 330.6 MB |
| AAUT_JVN29_S45_R1_001.fastq.gz | OTHER | 452.6 MB |
| AMPKInsInput_JVN21_S37_R1_001.fastq.gz | RNA-Seq | 373.0 MB |
| ampk_ins_input_JVN_70_S20_R1_001.fastq-2.gz | RNA-Seq | 356.1 MB |
| ampk_ins_input_JVN_70_S20_R1_001.fastq.gz | RNA-Seq | 126.1 MB |
| AMPKIns_JVN43_S59_R1_001.fastq.gz | OTHER | 433.6 MB |
| ampk_ins_JVN_59_S9_R1_001.fastq-2.gz | OTHER | 332.4 MB |
| ampk_ins_JVN_59_S9_R1_001.fastq.gz | OTHER | 57.4 MB |
| AMPKInsMetInput_JVN22_S38_R1_001.fastq.gz | RNA-Seq | 330.2 MB |
| ampk_ins_met_input_JVN_71_S21_R1_001.fastq-2.gz | RNA-Seq | 312.1 MB |
| ampk_ins_met_input_JVN_71_S21_R1_001.fastq.gz | RNA-Seq | 109.0 MB |
| AMPKInsMet_JVN44_S60_R1_001.fastq.gz | OTHER | 382.8 MB |
| ampk_ins_met_JVN_60_S10_R1_001.fastq-2.gz | OTHER | 374.9 MB |
| ampk_ins_met_JVN_60_S10_R1_001.fastq.gz | OTHER | 60.5 MB |
| AMPKMetInput_JVN11_S27_R1_001.fastq.gz | RNA-Seq | 390.5 MB |
| AMPKMet_JVN42_S58_R1_001.fastq.gz | OTHER | 414.0 MB |
| AMPKUTInput_JVN10_S26_R1_001.fastq.gz | RNA-Seq | 379.2 MB |
| AMPKUT_JVN41_S57_R1_001.fastq.gz | OTHER | 400.5 MB |
| TscInsInput_JVN17_S33_R1_001.fastq.gz | RNA-Seq | 373.7 MB |
| tsc_ins_input_JVN_66_S16_R1_001.fastq-2.gz | RNA-Seq | 381.5 MB |
| tsc_ins_input_JVN_66_S16_R1_001.fastq.gz | RNA-Seq | 131.6 MB |
| TscIns_JVN35_S51_R1_001.fastq.gz | OTHER | 357.4 MB |
| tsc_ins_JVN_55_S5_R1_001.fastq-2.gz | OTHER | 324.7 MB |
| tsc_ins_JVN_55_S5_R1_001.fastq.gz | OTHER | 138.9 MB |
| TscInsMetInput_JVN18_S34_R1_001.fastq.gz | RNA-Seq | 341.5 MB |
| tsc_ins_met_input_JVN_67_S17_R1_001.fastq-2.gz | RNA-Seq | 390.1 MB |
| tsc_ins_met_input_JVN_67_S17_R1_001.fastq.gz | RNA-Seq | 132.1 MB |
| TscInsMet_JVN36_S52_R1_001.fastq.gz | OTHER | 391.5 MB |
| tsc_ins_met_JVN_56_S6_R1_001.fastq-2.gz | OTHER | 365.0 MB |
| tsc_ins_met_JVN_56_S6_R1_001.fastq.gz | OTHER | 153.5 MB |
| TscMetinput_JVN7_S23_R1_001.fastq.gz | RNA-Seq | 337.1 MB |
| TscMet_JVN34_S50_R1_001.fastq.gz | OTHER | 391.1 MB |
| TscUTinput_JVN6_S22_R1_001.fastq.gz | RNA-Seq | 344.9 MB |
| TscUT_JVN33_S49_R1_001.fastq.gz | OTHER | 415.7 MB |
| WtInkinput_JVN3_S19_R1_001.fastq.gz | RNA-Seq | 336.4 MB |
| wt_ink_input_JVN_72_S22_R1_001.fastq-2.gz | RNA-Seq | 302.2 MB |
| wt_ink_input_JVN_72_S22_R1_001.fastq.gz | RNA-Seq | 107.1 MB |
| WtInk_JVN25_S41_R1_001.fastq.gz | OTHER | 419.7 MB |
| wt_ink_JVN_61_S11_R1_001.fastq-2.gz | OTHER | 378.1 MB |
| wt_ink_JVN_61_S11_R1_001.fastq.gz | OTHER | 64.9 MB |
| WTInsInkInput_JVN14_S30_R1_001.fastq.gz | RNA-Seq | 328.5 MB |
| WTInsInk_JVN28_S44_R1_001.fastq.gz | OTHER | 411.5 MB |
| WTInsInput_JVN12_S28_R1_001.fastq.gz | RNA-Seq | 395.5 MB |
| wt_ins_input_JVN_62_S12_R1_001.fastq-2.gz | RNA-Seq | 194.6 MB |
| wt_ins_input_JVN_62_S12_R1_001.fastq.gz | RNA-Seq | 127.8 MB |
| WTIns_JVN26_S42_R1_001.fastq.gz | OTHER | 391.5 MB |
| wt_ins_JVN_51_S1_R1_001.fastq-2.gz | OTHER | 449.5 MB |
| wt_ins_JVN_51_S1_R1_001.fastq.gz | OTHER | 195.5 MB |
| WTInsMetInput_JVN13_S29_R1_001.fastq.gz | RNA-Seq | 349.5 MB |
| wt_ins_met_input_JVN_63_S13_R1_001.fastq-2.gz | RNA-Seq | 338.2 MB |
| wt_ins_met_input_JVN_63_S13_R1_001.fastq.gz | RNA-Seq | 116.6 MB |
| WTInsMet_JVN27_S43_R1_001.fastq.gz | OTHER | 456.4 MB |
| wt_ins_met_JVN_52_S2_R1_001.fastq-2.gz | OTHER | 636.3 MB |
| wt_ins_met_JVN_52_S2_R1_001.fastq.gz | OTHER | 297.1 MB |
| WTMetinput_JVN2_S18_R1_001.fastq.gz | RNA-Seq | 336.6 MB |
| WTMet_JVN24_S40_R1_001.fastq.gz | OTHER | 385.5 MB |
| WTUTinput_JVN1_S17_R1_001.fastq.gz | RNA-Seq | 346.0 MB |
| WTUT_JVN23_S39_R1_001.fastq.gz | RNA-Seq | 401.5 MB |
Analysis Pipelines (4)
GSE157048 RNA-seq Data Processing
14 steps
GSE157049 RNA-seq Data Processing
14 steps
GSE157051 RNA-seq Data Processing
14 steps