title: SIRT3-dependent GOT2 acetylation status affects the malate-aspartate NADH shuttle activity and pancreatic tumor growth.
abstract: The malate-aspartate shuttle is indispensable for the net transfer of cytosolic NADH into mitochondria to maintain a high rate of glycolysis and to support rapid tumor cell growth. The malate-aspartate shuttle is operated by two pairs of enzymes that localize to the mitochondria and cytoplasm, glutamate oxaloacetate transaminases (GOT), and malate dehydrogenases (MDH). Here, we show that mitochondrial GOT2 is acetylated and that deacetylation depends on mitochondrial SIRT3. We have identified that acetylation occurs at three lysine residues, K159, K185, and K404 (3K), and enhances the association between GOT2 and MDH2. The GOT2 acetylation at these three residues promotes the net transfer of cytosolic NADH into mitochondria and changes the mitochondrial NADH/NAD(+) redox state to support ATP production. Additionally, GOT2 3K acetylation stimulates NADPH production to suppress ROS and to protect cells from oxidative damage. Moreover, GOT2 3K acetylation promotes pancreatic cell proliferation and tumor growth in vivo. Finally, we show that GOT2 K159 acetylation is increased in human pancreatic tumors, which correlates with reduced SIRT3 expression. Our study uncovers a previously unknown mechanism by which GOT2 acetylation stimulates the malate-aspartate NADH shuttle activity and oxidative protection.
authors.affiliation: State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Molecular and Cell Biology Lab, Institutes of Biomedical Sciences Shanghai Medical College Fudan University, Shanghai, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Molecular and Cell Biology Lab, Institutes of Biomedical Sciences Shanghai Medical College Fudan University, Shanghai, China.
authors.forename: Lisha
authors.initials: L
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Molecular and Cell Biology Lab, Institutes of Biomedical Sciences Shanghai Medical College Fudan University, Shanghai, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Shi
authors.affiliation: School of Pharmacy East China University of Science and Technology, Shanghai, China.
authors.forename: Yuzheng
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Molecular and Cell Biology Lab, Institutes of Biomedical Sciences Shanghai Medical College Fudan University, Shanghai, China.
authors.forename: Huaipeng
authors.initials: H
authors.lastname: Lin
authors.affiliation: State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Molecular and Cell Biology Lab, Institutes of Biomedical Sciences Shanghai Medical College Fudan University, Shanghai, China.
authors.forename: Mengli
authors.initials: M
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Molecular and Cell Biology Lab, Institutes of Biomedical Sciences Shanghai Medical College Fudan University, Shanghai, China.
authors.forename: Shimin
authors.initials: S
authors.lastname: Zhao
authors.affiliation: School of Pharmacy East China University of Science and Technology, Shanghai, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Zhejiang Cancer Research Institute Zhejiang Province Cancer Hospital Zhejiang Cancer Center, Hangzhou, China.
authors.forename: Zhi-Qiang
authors.initials: ZQ
authors.lastname: Ling
authors.affiliation: State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Molecular and Cell Biology Lab, Institutes of Biomedical Sciences Shanghai Medical College Fudan University, Shanghai, China Department of Pharmacology and Moores Cancer Center, University of California San Diego, La Jolla, CA, USA kuguan@ucsd.edu yxiong@email.unc.edu yedan@fudan.edu.cn.
authors.forename: Kun-Liang
authors.initials: KL
authors.lastname: Guan
authors.affiliation: State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Molecular and Cell Biology Lab, Institutes of Biomedical Sciences Shanghai Medical College Fudan University, Shanghai, China Lineberger Comprehensive Cancer Center, Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA kuguan@ucsd.edu yxiong@email.unc.edu yedan@fudan.edu.cn.
authors.forename: Yue
authors.initials: Y
authors.lastname: Xiong
authors.affiliation: State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Molecular and Cell Biology Lab, Institutes of Biomedical Sciences Shanghai Medical College Fudan University, Shanghai, China kuguan@ucsd.edu yxiong@email.unc.edu yedan@fudan.edu.cn.
authors.forename: Dan
authors.initials: D
authors.lastname: Ye
mesh_terms: D000107:Acetylation; D000818:Animals; D024442:Aspartate Aminotransferase, Mitochondrial / Q000378:metabolism*; D001224:Aspartic Acid / Q000378:metabolism*; D001692:Biological Transport; D021441:Carcinoma, Pancreatic Ductal / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D049109:Cell Proliferation / Q000235:genetics; D002478:Cells, Cultured; D057809:HEK293 Cells; D006801:Humans; D008293:Malates / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D009243:NAD / Q000378:metabolism; D010084:Oxidation-Reduction; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D011499:Protein Processing, Post-Translational / Q000502:physiology; D056566:Sirtuin 3 / Q000235:genetics / Q000378:metabolism*
chemical_list: D008293:Malates; D009243:NAD; D001224:Aspartic Acid; C030298:malic acid; D024442:Aspartate Aminotransferase, Mitochondrial; C466093:SIRT3 protein, human; D056566:Sirtuin 3
keywords: GOT2; acetylation; malate–aspartate NADH shuttle; pancreatic cancer