在酿酒酵母中研究人类mOGT基质导向序列的功能
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摘要
人类线粒体N-乙酰氨基葡萄糖转移酶(mitochondrial O-GlcNAc transferse,m OGT)通过其N端基质导向序列(matrix targeting sequence,MTS)定位于线粒体内膜上,m OGT的过表达将导致细胞的凋亡。蛋白质的O-Glc NAc修饰存在于绝大多数真核生物中,而酿酒酵母唯独缺乏该修饰途径。对人体m OGT过表达引起酿酒酵母生长缺陷的机制进行分析,期望利用该模型研究m OGT导致的细胞凋亡调控机制。在酿酒酵母细胞中表达人体m OGT及其截短序列发现:m OGT对酿酒酵母的生长抑制及细胞中的定位依赖于N端全长的MTS序列; MTS序列在酿酒酵母中共表达导致线粒体融合。MTS序列过表达的酿酒酵母菌可以作为研究m OGT引起细胞凋亡的模式细胞。
Human mitochondrial O-GlcNAc transferase (mOGT) localized in mitochondria inner membrane through its N-terminal martrix targeting sequence (MTS),mOGT overexpression caused cell spoptosis. O-GlcNAc modification existed in most eukaryote cells except Saccharomyces cerevisiae. In order to analyze mOGT induced mammalian cell apoptosis,the mechanism of mOGT caused yeast cell growth defect was investigated. Herein,mOGT was overexpressed in Saccharomyces cerevisiae. Both yeast growth defect and mOGT localization were largely depends on MTS sequence. Furthermore,MTS expression caused yeast mitochondria fussion. Therefore,MTS overexpressed yeast cells might be applied to analyze mOGT caused cell apoptosis.
Human mitochondrial O-GlcNAc transferase (mOGT) localized in mitochondria inner membrane through its N-terminal martrix targeting sequence (MTS),mOGT overexpression caused cell spoptosis. O-GlcNAc modification existed in most eukaryote cells except Saccharomyces cerevisiae. In order to analyze mOGT induced mammalian cell apoptosis,the mechanism of mOGT caused yeast cell growth defect was investigated. Herein,mOGT was overexpressed in Saccharomyces cerevisiae. Both yeast growth defect and mOGT localization were largely depends on MTS sequence. Furthermore,MTS expression caused yeast mitochondria fussion. Therefore,MTS overexpressed yeast cells might be applied to analyze mOGT caused cell apoptosis.
引文
[1]Hart G W.Minireview series on the thirtieth anniversary of research on O-Glc NAcylation of nuclear and cytoplasmic proteins:Nutrient regulation of cellular metabolism and physiology by O-Glc NAcylation.Journal of Biological Chemmistry,2014,289(50):34422-34423.
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[5]Marsh S A,Collins H E,Chatham J C.Protein O-Glc NAcylation and cardiovascular(patho)physiology.Journal of Biological Chemmistry,2014,289(50):34449-34456.
[6]Zhu Y,Shan X,Yuzwa S A,et al.The emerging link between O-Glc NAc and Alzheimer disease.Journal of Biological Chemmistry,2014,289(50):34472-34481.
[7]Hanover J A,Yu S,Lubas W B,et al.Mitochondrial and nucleocytoplasmic isoforms of O-linked Glc NAc transferase encoded by a single mammalian gene.Archives of Biochemistry Biophysics,2003,409(2):287-297.
[8]Burnham-Marusich A R,Berninsone P M.Multiple proteins with essential mitochondrial functions have glycosylated isoforms.Mitochondrion,2012,12(4):423-427.
[9]Ma J,Liu T,Wei A C,et al.O-Glc NAcomic profiling identifies widespread O-linkedβ-N-acetylglucosamine modification(O-Glc NAcylation)in oxidative phosphorylation system regulating cardiac mitochondrial function.Journal of Biological Chemmistry,2015,290(49):29141-29153.
[10]Hu Y,Suarez J,Fricovsky E,et al.Increased enzymatic O-Glc NAcylation of mitochondrial proteins impairs mitochondrial function in cardiac myocytes exposed to high glucose.Journal of Biological Chemmistry,2009,284(1):547-555.
[11]Lazarus M B,Nam Y,Jiang J,et al.Structure of human O-Glc NAc transferase and its complex with a peptide substrate.Nature,2011,469(7331):564-567.
[12]Sacoman J L,Dagda R Y,Burnham-Marusich A R,et al.Mitochondrial O-Glc NAc transferase(m OGT)regulates mitochondrial structure,function and survival in He La cells.Journal of Biological Chemmistry,2017,292(11):4499-4518.
[13]Halim A,Larsen I S,Neubert P,et al.Discovery of a nucleocytoplasmic O-mannose glycoproteome in yeast.Proceedings of the National Academy of Science of the United States of America,2015,112(51):15648-15653.
[14]Nakanishi H,Li F,Han B X,et al.Yeast cells as an assay system for in vivo O-Glc NAc modification.Biochimica et Biophysica acta-General Subjects,2017,1861(5):1159-1167.
[15]韩宝仙,高晓冬,中西秀树.人体线粒体N-乙酰氨基葡萄糖转移酶在酿酒酵母中的表达.食品与生物技术学报,2016,35(9):987-992.Han B X,Gao X D,Nakanishi H.Expression of a human mitochondrial N-acetylglucosamine transferase in Saccharomyces cerevisiae.Journal of Food Science and Biotechnology,2016,35(9):987-992.
[16]Longtine M S,Mckenzie Iii A,Demarini D J,et al.Additional modules for versatile and economical PCR‐based gene deletion and modification in Saccharomyces cerevisiae.Yeast,2010,14(10):953-961.
[17]Lubas W A,Frank D W,Krause M,et al.O-linked Glc NAc transferase is a conserved nucleocytoplasmic protein containing tetratricopeptide repeats.Journal of Biological Chemmistry,1997,272(14):9316-9324.
[18]Mumberg D,Müller R,Funk M.Yeast vectors for the controlled expression of heterologous proteins in different genetic backgrounds.Gene,1995,156(1):119-122.
[19]Banerjee P S,Ma J,Hart G W.Diabetes-associated dysregulation of O-Glc NAcylation in rat cardiac mitochondria.Proceedings of the National Academy of Science of the United States of America,2015,112(19):6050-6055.
[20]Love D C,Kochan J,Cathey R L,et al.Mitochondrial and nucleocytoplasmic targeting of O-linked Glc NAc transferase.Journal of Cell Science,2003,116(4):647-654.
[21]Shin S H,Love D C,Hanover J A.Elevated O-Glc NAcdependent signaling through inducible m OGT expression selectively triggers apoptosis.Amino Acids,2011,40(3):885-893.
[22]Bocharova N,Chave-Cox R,Sokolov S,et al.Protein aggregation and neurodegeneration:clues from a yeast model of Huntington’s disease.Biochemistry,2009,74(2):231-234.
[23]Zha H,Fisk H A,Yaffe M P,et al.Structure-function comparisons of the proapoptotic protein Bax in yeast and mammalian cells.Molecular and Cellular Biology,1996,16(11):6494-6508.
[24]Bossywetzel E,Barsoum M J,Godzik A,et al.Mitochondrial fission in apoptosis,neurodegeneration and aging.Current Opinion in Cell Biology,2003,15(6):706-716.
[25]Mignotte B,Vayssiere J L.Mitochondria and apoptosis.European Journal of Biochemistry,1998,252(1):1-15.
[26]安志远,丁文一.鲍曼不动杆菌Omp34经线粒体途径诱导He La细胞凋亡.生物技术,2018,28(4):323-328.An Z Y,Ding W Y.The outer membrane protein of Acinetobacter baumannii induces apoptosis in He La cells through mitochondria signal pathway.Biotechnology,2018,28(4):323-328.
[2]Marshall S,Bacote V,Traxinger R R.Discovery of a metabolic pathway mediating glucose-induced desensitization of the glucose transport system.Role of hexosamine biosynthesis in the induction of insulin resistance.Journal of Biological Chemmistry,1991,266(8):4706-4712.
[3]Vaidyanathan K,Wells L.Multiple tissue-specific roles for the O-Glc NAc post-translational modification in the induction of and complications arising from type II diabetes.Journal of Biological Chemmistry,2014,289(50):34466-34471.
[4]Ma Z,Vosseller K.Cancer metabolism and elevated O-Glc NAc in oncogenic signaling.Journal of Biological Chemmistry,2014,289(50):34457-34465.
[5]Marsh S A,Collins H E,Chatham J C.Protein O-Glc NAcylation and cardiovascular(patho)physiology.Journal of Biological Chemmistry,2014,289(50):34449-34456.
[6]Zhu Y,Shan X,Yuzwa S A,et al.The emerging link between O-Glc NAc and Alzheimer disease.Journal of Biological Chemmistry,2014,289(50):34472-34481.
[7]Hanover J A,Yu S,Lubas W B,et al.Mitochondrial and nucleocytoplasmic isoforms of O-linked Glc NAc transferase encoded by a single mammalian gene.Archives of Biochemistry Biophysics,2003,409(2):287-297.
[8]Burnham-Marusich A R,Berninsone P M.Multiple proteins with essential mitochondrial functions have glycosylated isoforms.Mitochondrion,2012,12(4):423-427.
[9]Ma J,Liu T,Wei A C,et al.O-Glc NAcomic profiling identifies widespread O-linkedβ-N-acetylglucosamine modification(O-Glc NAcylation)in oxidative phosphorylation system regulating cardiac mitochondrial function.Journal of Biological Chemmistry,2015,290(49):29141-29153.
[10]Hu Y,Suarez J,Fricovsky E,et al.Increased enzymatic O-Glc NAcylation of mitochondrial proteins impairs mitochondrial function in cardiac myocytes exposed to high glucose.Journal of Biological Chemmistry,2009,284(1):547-555.
[11]Lazarus M B,Nam Y,Jiang J,et al.Structure of human O-Glc NAc transferase and its complex with a peptide substrate.Nature,2011,469(7331):564-567.
[12]Sacoman J L,Dagda R Y,Burnham-Marusich A R,et al.Mitochondrial O-Glc NAc transferase(m OGT)regulates mitochondrial structure,function and survival in He La cells.Journal of Biological Chemmistry,2017,292(11):4499-4518.
[13]Halim A,Larsen I S,Neubert P,et al.Discovery of a nucleocytoplasmic O-mannose glycoproteome in yeast.Proceedings of the National Academy of Science of the United States of America,2015,112(51):15648-15653.
[14]Nakanishi H,Li F,Han B X,et al.Yeast cells as an assay system for in vivo O-Glc NAc modification.Biochimica et Biophysica acta-General Subjects,2017,1861(5):1159-1167.
[15]韩宝仙,高晓冬,中西秀树.人体线粒体N-乙酰氨基葡萄糖转移酶在酿酒酵母中的表达.食品与生物技术学报,2016,35(9):987-992.Han B X,Gao X D,Nakanishi H.Expression of a human mitochondrial N-acetylglucosamine transferase in Saccharomyces cerevisiae.Journal of Food Science and Biotechnology,2016,35(9):987-992.
[16]Longtine M S,Mckenzie Iii A,Demarini D J,et al.Additional modules for versatile and economical PCR‐based gene deletion and modification in Saccharomyces cerevisiae.Yeast,2010,14(10):953-961.
[17]Lubas W A,Frank D W,Krause M,et al.O-linked Glc NAc transferase is a conserved nucleocytoplasmic protein containing tetratricopeptide repeats.Journal of Biological Chemmistry,1997,272(14):9316-9324.
[18]Mumberg D,Müller R,Funk M.Yeast vectors for the controlled expression of heterologous proteins in different genetic backgrounds.Gene,1995,156(1):119-122.
[19]Banerjee P S,Ma J,Hart G W.Diabetes-associated dysregulation of O-Glc NAcylation in rat cardiac mitochondria.Proceedings of the National Academy of Science of the United States of America,2015,112(19):6050-6055.
[20]Love D C,Kochan J,Cathey R L,et al.Mitochondrial and nucleocytoplasmic targeting of O-linked Glc NAc transferase.Journal of Cell Science,2003,116(4):647-654.
[21]Shin S H,Love D C,Hanover J A.Elevated O-Glc NAcdependent signaling through inducible m OGT expression selectively triggers apoptosis.Amino Acids,2011,40(3):885-893.
[22]Bocharova N,Chave-Cox R,Sokolov S,et al.Protein aggregation and neurodegeneration:clues from a yeast model of Huntington’s disease.Biochemistry,2009,74(2):231-234.
[23]Zha H,Fisk H A,Yaffe M P,et al.Structure-function comparisons of the proapoptotic protein Bax in yeast and mammalian cells.Molecular and Cellular Biology,1996,16(11):6494-6508.
[24]Bossywetzel E,Barsoum M J,Godzik A,et al.Mitochondrial fission in apoptosis,neurodegeneration and aging.Current Opinion in Cell Biology,2003,15(6):706-716.
[25]Mignotte B,Vayssiere J L.Mitochondria and apoptosis.European Journal of Biochemistry,1998,252(1):1-15.
[26]安志远,丁文一.鲍曼不动杆菌Omp34经线粒体途径诱导He La细胞凋亡.生物技术,2018,28(4):323-328.An Z Y,Ding W Y.The outer membrane protein of Acinetobacter baumannii induces apoptosis in He La cells through mitochondria signal pathway.Biotechnology,2018,28(4):323-328.