#METABOLOMICS WORKBENCH yli_20210510_131741_mwtab.txt DATATRACK_ID:2632 STUDY_ID:ST001786 ANALYSIS_ID:AN002897 PROJECT_ID:PR001135 VERSION 1 CREATED_ON May 10, 2021, 2:41 pm #PROJECT PR:PROJECT_TITLE Multi-omics analysis of glucose-mediated signaling by a moonlighting Gβ protein PR:PROJECT_TITLE Asc1/RACK1 PR:PROJECT_TYPE C18 Reversed-Phase Broad Spectrum Metabolomics PR:PROJECT_SUMMARY While much is known about glucose metabolism in yeast, less is known about the PR:PROJECT_SUMMARY receptors and signaling pathways that indicate glucose availability. We obtained PR:PROJECT_SUMMARY metabolic profiles for wildtype and 16 mutants affecting the yeast glucose PR:PROJECT_SUMMARY sensing pathway, comparing 0.05% glucose vs 10 min after glucose addition to 2%. PR:PROJECT_SUMMARY First, we determined that the G protein-coupled receptor (Gpr1/Gpa2) directs PR:PROJECT_SUMMARY early events in glucose utilization while the transceptors (Snf3/Rgt2) regulate PR:PROJECT_SUMMARY subsequent processes and downstream products of glucose metabolism. Whereas the PR:PROJECT_SUMMARY large G protein transmits the signal from its cognate receptor, Ras2 (but not PR:PROJECT_SUMMARY Ras1) integrates responses from both receptor pathways. Second, we determined PR:PROJECT_SUMMARY the relative contributions of the G protein α (Gpa2) and β (Asc1) subunits to PR:PROJECT_SUMMARY glucose-initiated processes. We determined that Gpa2 is primarily involved in PR:PROJECT_SUMMARY regulating carbohydrate metabolism while Asc1 is primarily involved in amino PR:PROJECT_SUMMARY acid metabolism. Both proteins are involved in regulating purine metabolism. PR:PROJECT_SUMMARY Collectively, our analysis reveals the molecular basis for glucose detection and PR:PROJECT_SUMMARY the earliest events of glucose-dependent signal transduction in yeast. PR:INSTITUTE University of North Carolina at Chapel Hill PR:DEPARTMENT Nutrition PR:LABORATORY Metabolomics and Exposome Laboratory, Nutrition Research Institute, UNC Chapel PR:LABORATORY Hill PR:LAST_NAME Sumner PR:FIRST_NAME Susan PR:ADDRESS 500 Laureate Way, Nutrition Research Institute, UNC Chapel Hill PR:EMAIL susan_sumner@unc.edu PR:PHONE (704) 250-5066 #STUDY ST:STUDY_TITLE Multi-omics analysis of glucose-mediated signaling by a moonlighting Gβ protein ST:STUDY_TITLE Asc1/RACK1 ST:STUDY_TYPE Untargeted UPLC-MS Metabolomics Analysis ST:STUDY_SUMMARY While much is known about glucose metabolism in yeast, less is known about the ST:STUDY_SUMMARY receptors and signaling pathways that indicate glucose availability. We obtained ST:STUDY_SUMMARY metabolic profiles for wildtype and 16 mutants affecting the yeast glucose ST:STUDY_SUMMARY sensing pathway, comparing 0.05% glucose vs 10 min after glucose addition to 2%. ST:STUDY_SUMMARY First, we determined that the G protein-coupled receptor (Gpr1/Gpa2) directs ST:STUDY_SUMMARY early events in glucose utilization while the transceptors (Snf3/Rgt2) regulate ST:STUDY_SUMMARY subsequent processes and downstream products of glucose metabolism. Whereas the ST:STUDY_SUMMARY large G protein transmits the signal from its cognate receptor, Ras2 (but not ST:STUDY_SUMMARY Ras1) integrates responses from both receptor pathways. Second, we determined ST:STUDY_SUMMARY the relative contributions of the G protein α (Gpa2) and β (Asc1) subunits to ST:STUDY_SUMMARY glucose-initiated processes. We determined that Gpa2 is primarily involved in ST:STUDY_SUMMARY regulating carbohydrate metabolism while Asc1 is primarily involved in amino ST:STUDY_SUMMARY acid metabolism. Both proteins are involved in regulating purine metabolism. ST:STUDY_SUMMARY Collectively, our analysis reveals the molecular basis for glucose detection and ST:STUDY_SUMMARY the earliest events of glucose-dependent signal transduction in yeast. ST:INSTITUTE University of North Carolina at Chapel Hill ST:DEPARTMENT Nutrition ST:LABORATORY Metabolomics and Exposome Laboratory, Nutrition Research Institute, UNC Chapel ST:LABORATORY Hill ST:LAST_NAME Sumner ST:FIRST_NAME Susan ST:ADDRESS 500 Laureate Way, Nutrition Research Institute, UNC Chapel Hill ST:EMAIL susan_sumner@unc.edu ST:PHONE (919) 622-4456 ST:NUM_GROUPS 28 ST:TOTAL_SUBJECTS 192 #SUBJECT SU:SUBJECT_TYPE Yeast SU:SUBJECT_SPECIES Saccharomyces cerevisiae SU:GENDER Not applicable #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - asc1_H1_A Genotype:asc1 | Condition:H RAW_FILE_NAME=asc1_H1_A.RAW SUBJECT_SAMPLE_FACTORS - asc1_H3_A Genotype:asc1 | Condition:H RAW_FILE_NAME=asc1_H3_A.RAW SUBJECT_SAMPLE_FACTORS - asc1_H4_A Genotype:asc1 | Condition:H RAW_FILE_NAME=asc1_H4_A.RAW SUBJECT_SAMPLE_FACTORS - asc1_H5_A Genotype:asc1 | Condition:H RAW_FILE_NAME=asc1_H5_A.RAW SUBJECT_SAMPLE_FACTORS - asc1_H6_A Genotype:asc1 | Condition:H RAW_FILE_NAME=asc1_H6_A.RAW SUBJECT_SAMPLE_FACTORS - asc1_L1_A Genotype:asc1 | Condition:L RAW_FILE_NAME=asc1_L1_A.RAW SUBJECT_SAMPLE_FACTORS - asc1_L2_A Genotype:asc1 | Condition:L RAW_FILE_NAME=asc1_L2_A.RAW SUBJECT_SAMPLE_FACTORS - asc1_L3_A Genotype:asc1 | Condition:L RAW_FILE_NAME=asc1_L3_A.RAW SUBJECT_SAMPLE_FACTORS - asc1_L4_A Genotype:asc1 | Condition:L RAW_FILE_NAME=asc1_L4_A.RAW SUBJECT_SAMPLE_FACTORS - asc1_L5_A Genotype:asc1 | Condition:L RAW_FILE_NAME=asc1_L5_A.RAW SUBJECT_SAMPLE_FACTORS - asc1_L6_A Genotype:asc1 | Condition:L RAW_FILE_NAME=asc1_L6_A.RAW SUBJECT_SAMPLE_FACTORS - gpa2_H1_A Genotype:gpa2 | Condition:H RAW_FILE_NAME=gpa2_H1_A.RAW SUBJECT_SAMPLE_FACTORS - gpa2_H2_A Genotype:gpa2 | Condition:H RAW_FILE_NAME=gpa2_H2_A.RAW SUBJECT_SAMPLE_FACTORS - gpa2_H3_A Genotype:gpa2 | Condition:H RAW_FILE_NAME=gpa2_H3_A.RAW SUBJECT_SAMPLE_FACTORS - gpa2_H4_A Genotype:gpa2 | Condition:H RAW_FILE_NAME=gpa2_H4_A.RAW SUBJECT_SAMPLE_FACTORS - gpa2_H5_A Genotype:gpa2 | Condition:H RAW_FILE_NAME=gpa2_H5_A.RAW SUBJECT_SAMPLE_FACTORS - gpa2_H6_A Genotype:gpa2 | Condition:H RAW_FILE_NAME=gpa2_H6_A.RAW SUBJECT_SAMPLE_FACTORS - gpa2_L1_A Genotype:gpa2 | Condition:L RAW_FILE_NAME=gpa2_L1_A.RAW SUBJECT_SAMPLE_FACTORS - gpa2_L2_A Genotype:gpa2 | Condition:L RAW_FILE_NAME=gpa2_L2_A.RAW SUBJECT_SAMPLE_FACTORS - gpa2_L3_A Genotype:gpa2 | Condition:L RAW_FILE_NAME=gpa2_L3_A.RAW SUBJECT_SAMPLE_FACTORS - gpa2_L4_A Genotype:gpa2 | Condition:L RAW_FILE_NAME=gpa2_L4_A.RAW SUBJECT_SAMPLE_FACTORS - gpa2_L5_A Genotype:gpa2 | Condition:L RAW_FILE_NAME=gpa2_L5_A.RAW SUBJECT_SAMPLE_FACTORS - gpa2_L6_A Genotype:gpa2 | Condition:L RAW_FILE_NAME=gpa2_L6_A.RAW SUBJECT_SAMPLE_FACTORS - gpr1_H1_A Genotype:gpr1 | Condition:H RAW_FILE_NAME=gpr1_H1_A.RAW SUBJECT_SAMPLE_FACTORS - gpr1_H2_A Genotype:gpr1 | Condition:H RAW_FILE_NAME=gpr1_H2_A.RAW SUBJECT_SAMPLE_FACTORS - gpr1_H3_A Genotype:gpr1 | Condition:H RAW_FILE_NAME=gpr1_H3_A.RAW SUBJECT_SAMPLE_FACTORS - gpr1_H4_A Genotype:gpr1 | Condition:H RAW_FILE_NAME=gpr1_H4_A.RAW SUBJECT_SAMPLE_FACTORS - gpr1_H5_A Genotype:gpr1 | Condition:H RAW_FILE_NAME=gpr1_H5_A.RAW SUBJECT_SAMPLE_FACTORS - gpr1_H6_A Genotype:gpr1 | Condition:H RAW_FILE_NAME=gpr1_H6_A.RAW SUBJECT_SAMPLE_FACTORS - gpr1_L1_A Genotype:gpr1 | Condition:L RAW_FILE_NAME=gpr1_L1_A.RAW SUBJECT_SAMPLE_FACTORS - gpr1_L2_A Genotype:gpr1 | Condition:L RAW_FILE_NAME=gpr1_L2_A.RAW SUBJECT_SAMPLE_FACTORS - gpr1_L3_A Genotype:gpr1 | Condition:L RAW_FILE_NAME=gpr1_L3_A.RAW SUBJECT_SAMPLE_FACTORS - gpr1_L4_A Genotype:gpr1 | Condition:L RAW_FILE_NAME=gpr1_L4_A.RAW SUBJECT_SAMPLE_FACTORS - gpr1_L5_A Genotype:gpr1 | Condition:L RAW_FILE_NAME=gpr1_L5_A.RAW SUBJECT_SAMPLE_FACTORS - gpr1_L6_A Genotype:gpr1 | Condition:L RAW_FILE_NAME=gpr1_L6_A.RAW SUBJECT_SAMPLE_FACTORS - pde1_H1_B Genotype:pde1 | Condition:H RAW_FILE_NAME=pde1_H1_B.RAW SUBJECT_SAMPLE_FACTORS - pde1_H2_B Genotype:pde1 | Condition:H RAW_FILE_NAME=pde1_H2_B.RAW SUBJECT_SAMPLE_FACTORS - pde1_H3_B Genotype:pde1 | Condition:H RAW_FILE_NAME=pde1_H3_B.RAW SUBJECT_SAMPLE_FACTORS - pde1_H4_B Genotype:pde1 | Condition:H RAW_FILE_NAME=pde1_H4_B.RAW SUBJECT_SAMPLE_FACTORS - pde1_H5_B Genotype:pde1 | Condition:H RAW_FILE_NAME=pde1_H5_B.RAW SUBJECT_SAMPLE_FACTORS - pde1_H6_B Genotype:pde1 | Condition:H RAW_FILE_NAME=pde1_H6_B.RAW SUBJECT_SAMPLE_FACTORS - pde1_L1_B Genotype:pde1 | Condition:L RAW_FILE_NAME=pde1_L1_B.RAW SUBJECT_SAMPLE_FACTORS - pde1_L2_B Genotype:pde1 | Condition:L RAW_FILE_NAME=pde1_L2_B.RAW SUBJECT_SAMPLE_FACTORS - pde1_L3_B Genotype:pde1 | Condition:L RAW_FILE_NAME=pde1_L3_B.RAW SUBJECT_SAMPLE_FACTORS - pde1_L4_B Genotype:pde1 | Condition:L RAW_FILE_NAME=pde1_L4_B.RAW SUBJECT_SAMPLE_FACTORS - pde1_L5_B Genotype:pde1 | Condition:L RAW_FILE_NAME=pde1_L5_B.RAW SUBJECT_SAMPLE_FACTORS - pde1_L6_B Genotype:pde1 | Condition:L RAW_FILE_NAME=pde1_L6_B.RAW SUBJECT_SAMPLE_FACTORS - pde2_H1_B Genotype:pde2 | Condition:H RAW_FILE_NAME=pde2_H1_B.RAW SUBJECT_SAMPLE_FACTORS - pde2_H2_B Genotype:pde2 | Condition:H RAW_FILE_NAME=pde2_H2_B.RAW SUBJECT_SAMPLE_FACTORS - pde2_H3_B Genotype:pde2 | Condition:H RAW_FILE_NAME=pde2_H3_B.RAW SUBJECT_SAMPLE_FACTORS - pde2_H4_B Genotype:pde2 | Condition:H RAW_FILE_NAME=pde2_H4_B.RAW SUBJECT_SAMPLE_FACTORS - pde2_H5_B Genotype:pde2 | Condition:H RAW_FILE_NAME=pde2_H5_B.RAW SUBJECT_SAMPLE_FACTORS - pde2_H6_B Genotype:pde2 | Condition:H RAW_FILE_NAME=pde2_H6_B.RAW SUBJECT_SAMPLE_FACTORS - pde2_L1_B Genotype:pde2 | Condition:L RAW_FILE_NAME=pde2_L1_B.RAW SUBJECT_SAMPLE_FACTORS - pde2_L2_B Genotype:pde2 | Condition:L RAW_FILE_NAME=pde2_L2_B.RAW SUBJECT_SAMPLE_FACTORS - pde2_L3_B Genotype:pde2 | Condition:L RAW_FILE_NAME=pde2_L3_B.RAW SUBJECT_SAMPLE_FACTORS - pde2_L4_B Genotype:pde2 | Condition:L RAW_FILE_NAME=pde2_L4_B.RAW SUBJECT_SAMPLE_FACTORS - pde2_L5_B Genotype:pde2 | Condition:L RAW_FILE_NAME=pde2_L5_B.RAW SUBJECT_SAMPLE_FACTORS - pde2_L6_B Genotype:pde2 | Condition:L RAW_FILE_NAME=pde2_L6_B.RAW SUBJECT_SAMPLE_FACTORS - ras1_H1_B Genotype:ras1 | Condition:H RAW_FILE_NAME=ras1_H1_B.RAW SUBJECT_SAMPLE_FACTORS - ras1_H2_B Genotype:ras1 | Condition:H RAW_FILE_NAME=ras1_H2_B.RAW SUBJECT_SAMPLE_FACTORS - ras1_H3_B Genotype:ras1 | Condition:H RAW_FILE_NAME=ras1_H3_B.RAW SUBJECT_SAMPLE_FACTORS - ras1_H4_B Genotype:ras1 | Condition:H RAW_FILE_NAME=ras1_H4_B.RAW SUBJECT_SAMPLE_FACTORS - ras1_H5_B Genotype:ras1 | Condition:H RAW_FILE_NAME=ras1_H5_B.RAW SUBJECT_SAMPLE_FACTORS - ras1_H6_B Genotype:ras1 | Condition:H RAW_FILE_NAME=ras1_H6_B.RAW SUBJECT_SAMPLE_FACTORS - ras1_L1_B Genotype:ras1 | Condition:L RAW_FILE_NAME=ras1_L1_B.RAW SUBJECT_SAMPLE_FACTORS - ras1_L2_B Genotype:ras1 | Condition:L RAW_FILE_NAME=ras1_L2_B.RAW SUBJECT_SAMPLE_FACTORS - ras1_L3_B Genotype:ras1 | Condition:L RAW_FILE_NAME=ras1_L3_B.RAW SUBJECT_SAMPLE_FACTORS - ras1_L4_B Genotype:ras1 | Condition:L RAW_FILE_NAME=ras1_L4_B.RAW SUBJECT_SAMPLE_FACTORS - ras1_L5_B Genotype:ras1 | Condition:L RAW_FILE_NAME=ras1_L5_B.RAW SUBJECT_SAMPLE_FACTORS - ras1_L6_B Genotype:ras1 | Condition:L RAW_FILE_NAME=ras1_L6_B.RAW SUBJECT_SAMPLE_FACTORS - ras2_H1_B Genotype:ras2 | Condition:H RAW_FILE_NAME=ras2_H1_B.RAW SUBJECT_SAMPLE_FACTORS - ras2_H2_B Genotype:ras2 | Condition:H RAW_FILE_NAME=ras2_H2_B.RAW SUBJECT_SAMPLE_FACTORS - ras2_H3_B Genotype:ras2 | Condition:H RAW_FILE_NAME=ras2_H3_B.RAW SUBJECT_SAMPLE_FACTORS - ras2_H4_B Genotype:ras2 | Condition:H RAW_FILE_NAME=ras2_H4_B.RAW SUBJECT_SAMPLE_FACTORS - ras2_H5_B Genotype:ras2 | Condition:H RAW_FILE_NAME=ras2_H5_B.RAW SUBJECT_SAMPLE_FACTORS - ras2_H6_B Genotype:ras2 | Condition:H RAW_FILE_NAME=ras2_H6_B.RAW SUBJECT_SAMPLE_FACTORS - ras2_L1_B Genotype:ras2 | Condition:L RAW_FILE_NAME=ras2_L1_B.RAW SUBJECT_SAMPLE_FACTORS - ras2_L2_B Genotype:ras2 | Condition:L RAW_FILE_NAME=ras2_L2_B.RAW SUBJECT_SAMPLE_FACTORS - ras2_L3_B Genotype:ras2 | Condition:L RAW_FILE_NAME=ras2_L3_B.RAW SUBJECT_SAMPLE_FACTORS - ras2_L4_B Genotype:ras2 | Condition:L RAW_FILE_NAME=ras2_L4_B.RAW SUBJECT_SAMPLE_FACTORS - ras2_L5_B Genotype:ras2 | Condition:L RAW_FILE_NAME=ras2_L5_B.RAW SUBJECT_SAMPLE_FACTORS - ras2_L6_B Genotype:ras2 | Condition:L RAW_FILE_NAME=ras2_L6_B.RAW SUBJECT_SAMPLE_FACTORS - rgs2_H1_A Genotype:rgs2 | Condition:H RAW_FILE_NAME=rgs2_H1_A.RAW SUBJECT_SAMPLE_FACTORS - rgs2_H2_A Genotype:rgs2 | Condition:H RAW_FILE_NAME=rgs2_H2_A.RAW SUBJECT_SAMPLE_FACTORS - rgs2_H3_A Genotype:rgs2 | Condition:H RAW_FILE_NAME=rgs2_H3_A.RAW SUBJECT_SAMPLE_FACTORS - rgs2_H4_A Genotype:rgs2 | Condition:H RAW_FILE_NAME=rgs2_H4_A.RAW SUBJECT_SAMPLE_FACTORS - rgs2_H5_A Genotype:rgs2 | Condition:H RAW_FILE_NAME=rgs2_H5_A.RAW SUBJECT_SAMPLE_FACTORS - rgs2_H6_A Genotype:rgs2 | Condition:H RAW_FILE_NAME=rgs2_H6_A.RAW SUBJECT_SAMPLE_FACTORS - rgs2_L1_A Genotype:rgs2 | Condition:L RAW_FILE_NAME=rgs2_L1_A.RAW SUBJECT_SAMPLE_FACTORS - rgs2_L2_A Genotype:rgs2 | Condition:L RAW_FILE_NAME=rgs2_L2_A.RAW SUBJECT_SAMPLE_FACTORS - rgs2_L3_A Genotype:rgs2 | Condition:L RAW_FILE_NAME=rgs2_L3_A.RAW SUBJECT_SAMPLE_FACTORS - rgs2_L4_A Genotype:rgs2 | Condition:L RAW_FILE_NAME=rgs2_L4_A.RAW SUBJECT_SAMPLE_FACTORS - rgs2_L5_A Genotype:rgs2 | Condition:L RAW_FILE_NAME=rgs2_L5_A.RAW SUBJECT_SAMPLE_FACTORS - rgs2_L6_A Genotype:rgs2 | Condition:L RAW_FILE_NAME=rgs2_L6_A.RAW SUBJECT_SAMPLE_FACTORS - rgt2_H1_C Genotype:rgt2 | Condition:H RAW_FILE_NAME=rgt2_H1_C.RAW SUBJECT_SAMPLE_FACTORS - rgt2_H2_C Genotype:rgt2 | Condition:H RAW_FILE_NAME=rgt2_H2_C.RAW SUBJECT_SAMPLE_FACTORS - rgt2_H3_C Genotype:rgt2 | Condition:H RAW_FILE_NAME=rgt2_H3_C.RAW SUBJECT_SAMPLE_FACTORS - rgt2_H4_C Genotype:rgt2 | Condition:H RAW_FILE_NAME=rgt2_H4_C.RAW SUBJECT_SAMPLE_FACTORS - rgt2_H5_C Genotype:rgt2 | Condition:H RAW_FILE_NAME=rgt2_H5_C.RAW SUBJECT_SAMPLE_FACTORS - rgt2_H6_C Genotype:rgt2 | Condition:H RAW_FILE_NAME=rgt2_H6_C.RAW SUBJECT_SAMPLE_FACTORS - rgt2_L1_C Genotype:rgt2 | Condition:L RAW_FILE_NAME=rgt2_L1_C.RAW SUBJECT_SAMPLE_FACTORS - rgt2_L2_C Genotype:rgt2 | Condition:L RAW_FILE_NAME=rgt2_L2_C.RAW SUBJECT_SAMPLE_FACTORS - rgt2_L3_C Genotype:rgt2 | Condition:L RAW_FILE_NAME=rgt2_L3_C.RAW SUBJECT_SAMPLE_FACTORS - rgt2_L4_C Genotype:rgt2 | Condition:L RAW_FILE_NAME=rgt2_L4_C.RAW SUBJECT_SAMPLE_FACTORS - rgt2_L5_C Genotype:rgt2 | Condition:L RAW_FILE_NAME=rgt2_L5_C.RAW SUBJECT_SAMPLE_FACTORS - rgt2_L6_C Genotype:rgt2 | Condition:L RAW_FILE_NAME=rgt2_L6_C.RAW SUBJECT_SAMPLE_FACTORS - snf3_H1_C Genotype:snf3 | Condition:H RAW_FILE_NAME=snf3_H1_C.RAW SUBJECT_SAMPLE_FACTORS - snf3_H2_C Genotype:snf3 | Condition:H RAW_FILE_NAME=snf3_H2_C.RAW SUBJECT_SAMPLE_FACTORS - snf3_H3_C Genotype:snf3 | Condition:H RAW_FILE_NAME=snf3_H3_C.RAW SUBJECT_SAMPLE_FACTORS - snf3_H4_C Genotype:snf3 | Condition:H RAW_FILE_NAME=snf3_H4_C.RAW SUBJECT_SAMPLE_FACTORS - snf3_H5_C Genotype:snf3 | Condition:H RAW_FILE_NAME=snf3_H5_C.RAW SUBJECT_SAMPLE_FACTORS - snf3_H6_C Genotype:snf3 | Condition:H RAW_FILE_NAME=snf3_H6_C.RAW SUBJECT_SAMPLE_FACTORS - snf3_L1_C Genotype:snf3 | Condition:L RAW_FILE_NAME=snf3_L1_C.RAW SUBJECT_SAMPLE_FACTORS - snf3_L2_C Genotype:snf3 | Condition:L RAW_FILE_NAME=snf3_L2_C.RAW SUBJECT_SAMPLE_FACTORS - snf3_L3_C Genotype:snf3 | Condition:L RAW_FILE_NAME=snf3_L3_C.RAW SUBJECT_SAMPLE_FACTORS - snf3_L4_C Genotype:snf3 | Condition:L RAW_FILE_NAME=snf3_L4_C.RAW SUBJECT_SAMPLE_FACTORS - snf3_L5_C Genotype:snf3 | Condition:L RAW_FILE_NAME=snf3_L5_C.RAW SUBJECT_SAMPLE_FACTORS - snf3_L6_C Genotype:snf3 | Condition:L RAW_FILE_NAME=snf3_L6_C.RAW SUBJECT_SAMPLE_FACTORS - snf3rgt2_H1_C Genotype:snf3rgt2 | Condition:H RAW_FILE_NAME=snf3rgt2_H1_C.RAW SUBJECT_SAMPLE_FACTORS - snf3rgt2_H2_C Genotype:snf3rgt2 | Condition:H RAW_FILE_NAME=snf3rgt2_H2_C.RAW SUBJECT_SAMPLE_FACTORS - snf3rgt2_H3_C Genotype:snf3rgt2 | Condition:H RAW_FILE_NAME=snf3rgt2_H3_C.RAW SUBJECT_SAMPLE_FACTORS - snf3rgt2_H4_C Genotype:snf3rgt2 | Condition:H RAW_FILE_NAME=snf3rgt2_H4_C.RAW SUBJECT_SAMPLE_FACTORS - snf3rgt2_H5_C Genotype:snf3rgt2 | Condition:H RAW_FILE_NAME=snf3rgt2_H5_C.RAW SUBJECT_SAMPLE_FACTORS - snf3rgt2_H6_C Genotype:snf3rgt2 | Condition:H RAW_FILE_NAME=snf3rgt2_H6_C.RAW SUBJECT_SAMPLE_FACTORS - snf3rgt2_L1_C Genotype:snf3rgt2 | Condition:L RAW_FILE_NAME=snf3rgt2_L1_C.RAW SUBJECT_SAMPLE_FACTORS - snf3rgt2_L2_C Genotype:snf3rgt2 | Condition:L RAW_FILE_NAME=snf3rgt2_L2_C.RAW SUBJECT_SAMPLE_FACTORS - snf3rgt2_L3_C Genotype:snf3rgt2 | Condition:L RAW_FILE_NAME=snf3rgt2_L3_C.RAW SUBJECT_SAMPLE_FACTORS - snf3rgt2_L4_C Genotype:snf3rgt2 | Condition:L RAW_FILE_NAME=snf3rgt2_L4_C.RAW SUBJECT_SAMPLE_FACTORS - snf3rgt2_L5_C Genotype:snf3rgt2 | Condition:L RAW_FILE_NAME=snf3rgt2_L5_C.RAW SUBJECT_SAMPLE_FACTORS - snf3rgt2_L6_C Genotype:snf3rgt2 | Condition:L RAW_FILE_NAME=snf3rgt2_L6_C.RAW SUBJECT_SAMPLE_FACTORS - WT_H1_A Genotype:WT | Condition:H RAW_FILE_NAME=WT_H1_A.RAW SUBJECT_SAMPLE_FACTORS - WT_H1_B Genotype:WT | Condition:H RAW_FILE_NAME=WT_H1_B.RAW SUBJECT_SAMPLE_FACTORS - WT_H1_C Genotype:WT | Condition:H RAW_FILE_NAME=WT_H1_C.RAW SUBJECT_SAMPLE_FACTORS - WT_H2_A Genotype:WT | Condition:H RAW_FILE_NAME=WT_H2_A.RAW SUBJECT_SAMPLE_FACTORS - WT_H2_B Genotype:WT | Condition:H RAW_FILE_NAME=WT_H2_B.RAW SUBJECT_SAMPLE_FACTORS - WT_H2_C Genotype:WT | Condition:H RAW_FILE_NAME=WT_H2_C.RAW SUBJECT_SAMPLE_FACTORS - WT_H3_A Genotype:WT | Condition:H RAW_FILE_NAME=WT_H3_A.RAW SUBJECT_SAMPLE_FACTORS - WT_H3_B Genotype:WT | Condition:H RAW_FILE_NAME=WT_H3_B.RAW SUBJECT_SAMPLE_FACTORS - WT_H3_C Genotype:WT | Condition:H RAW_FILE_NAME=WT_H3_C.RAW SUBJECT_SAMPLE_FACTORS - WT_H4_A Genotype:WT | Condition:H RAW_FILE_NAME=WT_H4_A.RAW SUBJECT_SAMPLE_FACTORS - WT_H4_B Genotype:WT | Condition:H RAW_FILE_NAME=WT_H4_B.RAW SUBJECT_SAMPLE_FACTORS - WT_H4_C Genotype:WT | Condition:H RAW_FILE_NAME=WT_H4_C.RAW SUBJECT_SAMPLE_FACTORS - WT_H5_A Genotype:WT | Condition:H RAW_FILE_NAME=WT_H5_A.RAW SUBJECT_SAMPLE_FACTORS - WT_H5_B Genotype:WT | Condition:H RAW_FILE_NAME=WT_H5_B.RAW SUBJECT_SAMPLE_FACTORS - WT_H5_C Genotype:WT | Condition:H RAW_FILE_NAME=WT_H5_C.RAW SUBJECT_SAMPLE_FACTORS - WT_H6_A Genotype:WT | Condition:H RAW_FILE_NAME=WT_H6_A.RAW SUBJECT_SAMPLE_FACTORS - WT_H6_B Genotype:WT | Condition:H RAW_FILE_NAME=WT_H6_B.RAW SUBJECT_SAMPLE_FACTORS - WT_H6_C Genotype:WT | Condition:H RAW_FILE_NAME=WT_H6_C.RAW SUBJECT_SAMPLE_FACTORS - WT_L1_A Genotype:WT | Condition:L RAW_FILE_NAME=WT_L1_A.RAW SUBJECT_SAMPLE_FACTORS - WT_L1_B Genotype:WT | Condition:L RAW_FILE_NAME=WT_L1_B.RAW SUBJECT_SAMPLE_FACTORS - WT_L1_C Genotype:WT | Condition:L RAW_FILE_NAME=WT_L1_C.RAW SUBJECT_SAMPLE_FACTORS - WT_L2_A Genotype:WT | Condition:L RAW_FILE_NAME=WT_L2_A.RAW SUBJECT_SAMPLE_FACTORS - WT_L2_B Genotype:WT | Condition:L RAW_FILE_NAME=WT_L2_B.RAW SUBJECT_SAMPLE_FACTORS - WT_L2_C Genotype:WT | Condition:L RAW_FILE_NAME=WT_L2_C.RAW SUBJECT_SAMPLE_FACTORS - WT_L3_A Genotype:WT | Condition:L RAW_FILE_NAME=WT_L3_A.RAW SUBJECT_SAMPLE_FACTORS - WT_L3_B Genotype:WT | Condition:L RAW_FILE_NAME=WT_L3_B.RAW SUBJECT_SAMPLE_FACTORS - WT_L3_C Genotype:WT | Condition:L RAW_FILE_NAME=WT_L3_C.RAW SUBJECT_SAMPLE_FACTORS - WT_L4_A Genotype:WT | Condition:L RAW_FILE_NAME=WT_L4_A.RAW SUBJECT_SAMPLE_FACTORS - WT_L4_B Genotype:WT | Condition:L RAW_FILE_NAME=WT_L4_B.RAW SUBJECT_SAMPLE_FACTORS - WT_L4_C Genotype:WT | Condition:L RAW_FILE_NAME=WT_L4_C.RAW SUBJECT_SAMPLE_FACTORS - WT_L5_A Genotype:WT | Condition:L RAW_FILE_NAME=WT_L5_A.RAW SUBJECT_SAMPLE_FACTORS - WT_L5_B Genotype:WT | Condition:L RAW_FILE_NAME=WT_L5_B.RAW SUBJECT_SAMPLE_FACTORS - WT_L5_C Genotype:WT | Condition:L RAW_FILE_NAME=WT_L5_C.RAW SUBJECT_SAMPLE_FACTORS - WT_L6_A Genotype:WT | Condition:L RAW_FILE_NAME=WT_L6_A.RAW SUBJECT_SAMPLE_FACTORS - WT_L6_B Genotype:WT | Condition:L RAW_FILE_NAME=WT_L6_B.RAW SUBJECT_SAMPLE_FACTORS - WT_L6_C Genotype:WT | Condition:L RAW_FILE_NAME=WT_L6_C.RAW SUBJECT_SAMPLE_FACTORS - yck1_H1_C Genotype:yck1 | Condition:H RAW_FILE_NAME=yck1_H1_C.RAW SUBJECT_SAMPLE_FACTORS - yck1_H2_C Genotype:yck1 | Condition:H RAW_FILE_NAME=yck1_H2_C.RAW SUBJECT_SAMPLE_FACTORS - yck1_H3_C Genotype:yck1 | Condition:H RAW_FILE_NAME=yck1_H3_C.RAW SUBJECT_SAMPLE_FACTORS - yck1_H4_C Genotype:yck1 | Condition:H RAW_FILE_NAME=yck1_H4_C.RAW SUBJECT_SAMPLE_FACTORS - yck1_H5_C Genotype:yck1 | Condition:H RAW_FILE_NAME=yck1_H5_C.RAW SUBJECT_SAMPLE_FACTORS - yck1_H6_C Genotype:yck1 | Condition:H RAW_FILE_NAME=yck1_H6_C.RAW SUBJECT_SAMPLE_FACTORS - yck1_L1_C Genotype:yck1 | Condition:L RAW_FILE_NAME=yck1_L1_C.RAW SUBJECT_SAMPLE_FACTORS - yck1_L2_C Genotype:yck1 | Condition:L RAW_FILE_NAME=yck1_L2_C.RAW SUBJECT_SAMPLE_FACTORS - yck1_L3_C Genotype:yck1 | Condition:L RAW_FILE_NAME=yck1_L3_C.RAW SUBJECT_SAMPLE_FACTORS - yck1_L4_C Genotype:yck1 | Condition:L RAW_FILE_NAME=yck1_L4_C.RAW SUBJECT_SAMPLE_FACTORS - yck1_L5_C Genotype:yck1 | Condition:L RAW_FILE_NAME=yck1_L5_C.RAW SUBJECT_SAMPLE_FACTORS - yck1_L6_C Genotype:yck1 | Condition:L RAW_FILE_NAME=yck1_L6_C.RAW SUBJECT_SAMPLE_FACTORS - yck2_H1_C Genotype:yck2 | Condition:H RAW_FILE_NAME=yck2_H1_C.RAW SUBJECT_SAMPLE_FACTORS - yck2_H2_C Genotype:yck2 | Condition:H RAW_FILE_NAME=yck2_H2_C.RAW SUBJECT_SAMPLE_FACTORS - yck2_H3_C Genotype:yck2 | Condition:H RAW_FILE_NAME=yck2_H3_C.RAW SUBJECT_SAMPLE_FACTORS - yck2_H4_C Genotype:yck2 | Condition:H RAW_FILE_NAME=yck2_H4_C.RAW SUBJECT_SAMPLE_FACTORS - yck2_H5_C Genotype:yck2 | Condition:H RAW_FILE_NAME=yck2_H5_C.RAW SUBJECT_SAMPLE_FACTORS - yck2_H6_C Genotype:yck2 | Condition:H RAW_FILE_NAME=yck2_H6_C.RAW SUBJECT_SAMPLE_FACTORS - yck2_L1_C Genotype:yck2 | Condition:L RAW_FILE_NAME=yck2_L1_C.RAW SUBJECT_SAMPLE_FACTORS - yck2_L2_C Genotype:yck2 | Condition:L RAW_FILE_NAME=yck2_L2_C.RAW SUBJECT_SAMPLE_FACTORS - yck2_L3_C Genotype:yck2 | Condition:L RAW_FILE_NAME=yck2_L3_C.RAW SUBJECT_SAMPLE_FACTORS - yck2_L4_C Genotype:yck2 | Condition:L RAW_FILE_NAME=yck2_L4_C.RAW SUBJECT_SAMPLE_FACTORS - yck2_L5_C Genotype:yck2 | Condition:L RAW_FILE_NAME=yck2_L5_C.RAW SUBJECT_SAMPLE_FACTORS - yck2_L6_C Genotype:yck2 | Condition:L RAW_FILE_NAME=yck2_L6_C.RAW #COLLECTION CO:COLLECTION_SUMMARY For metabolomics, each replicate consisting of 3 mL of cell culture was mixed CO:COLLECTION_SUMMARY with 45 mL cold pure methanol on dry ice. After 5 min, cells were centrifuged in CO:COLLECTION_SUMMARY a precooled rotor (-80 °C). After discarding the supernatant, cell pellets were CO:COLLECTION_SUMMARY immediately stored at -80 °C. A small aliquot of each sample was saved to CO:COLLECTION_SUMMARY manually determine cell density with a hemocytometer. CO:SAMPLE_TYPE Yeast cells CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY No treatment in this study. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Frozen cell pellets were resuspended with extraction reagent (8:2 methanol-water SP:SAMPLEPREP_SUMMARY solution) to 3x108 cell/mL and then transferred into 2 mL ceramic bead SP:SAMPLEPREP_SUMMARY MagNalyser tubes. Blank samples were prepared by adding 1300 μL of extraction SP:SAMPLEPREP_SUMMARY reagent with no cells to a MagNalyser tube with ceramic beads. Tubes were SP:SAMPLEPREP_SUMMARY subjected to homogenization, with Bead Ruptor Elite Bead Mill Homogenizer (OMNI SP:SAMPLEPREP_SUMMARY International) at 6.0 m/s for 40 sec in 2 cycles at room temperature. This step SP:SAMPLEPREP_SUMMARY was repeated twice. All samples were then centrifuged at 16,000 xg for 10 min at SP:SAMPLEPREP_SUMMARY 4 °C. 500 μL of the supernatant was transferred into low-bind 1.7 mL microfuge SP:SAMPLEPREP_SUMMARY tubes. Total pools were made by combining an additional 65 μL of the SP:SAMPLEPREP_SUMMARY supernatant from each sample and then aliquoting this mixture into low-bind 1.7 SP:SAMPLEPREP_SUMMARY mL tubes at a volume of 500 μL. The remaining supernatant was stored at -80 °C SP:SAMPLEPREP_SUMMARY for repeat experiments if necessary. For all experimental samples, pooled SP:SAMPLEPREP_SUMMARY samples and blanks were dried using a speedvac vacuum concentrator overnight. SP:SAMPLEPREP_SUMMARY Dried samples were stored at -80 °C. Before LC-MS analysis, 100 μL of SP:SAMPLEPREP_SUMMARY reconstitution buffer (95:5 water:methanol with 500 ng/mL tryptophan d-5) was SP:SAMPLEPREP_SUMMARY added to each dried sample. All tubes were vortexed at 5000 rpm for 10 min and SP:SAMPLEPREP_SUMMARY then centrifuged at room temperature at 16,000 xg for 4 min. Supernatant was SP:SAMPLEPREP_SUMMARY transferred into autosampler vials for LC-MS. SP:PROCESSING_STORAGE_CONDITIONS 4℃ SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY None CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Waters Acquity BEH HSS T3 (100 x 2.1mm, 1.8um) CH:FLOW_RATE 0.4 mL/min CH:COLUMN_TEMPERATURE 8 CH:SOLVENT_A Water with 0.1% Formic Acid CH:SOLVENT_B Methanol with 0.1% Formic Acid CH:COLUMN_PRESSURE 6000-10000 CH:INJECTION_TEMPERATURE 8 CH:WEAK_WASH_SOLVENT_NAME 10:90 Methanol:Water with 0.1% FA solution CH:STRONG_WASH_SOLVENT_NAME 75:25 2-Propanol: Water with 0.1% FA solution CH:RANDOMIZATION_ORDER Yes CH:CHROMATOGRAPHY_COMMENTS Time(min) Flow Rate %A %B Curve 1. 0 0.4 99.0 1.0 5 2. 1.00 0.4 99.0 1.0 5 3. CH:CHROMATOGRAPHY_COMMENTS 16.00 0.4 1.0 99.0 5 4. 19.00 0.4 1.0 99.0 5 5. 19.50 0.4 99.0 1.0 5 #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME UNC-NRI Sumner lab AN:OPERATOR_NAME Yuanyuan Li & Blake Rushing AN:DETECTOR_TYPE Orbitrap AN:SOFTWARE_VERSION Xcalibur 4.1 AN:DATA_FORMAT profile #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF hybrid Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS NA MS:CAPILLARY_TEMPERATURE 275 °C MS:CAPILLARY_VOLTAGE 3.5 KV MS:COLLISION_ENERGY 10-35, ramp MS:COLLISION_GAS N2 MS:DRY_GAS_FLOW 45 MS:DRY_GAS_TEMP 325°C MS:FRAGMENTATION_METHOD CID MS:DESOLVATION_GAS_FLOW 45 MS:MS_RESULTS_FILE ST001786_AN002897_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes #END