2000 Fiscal Year Annual Research Report

糖鎖遺伝子による細胞リモデリング

Research Project

Project/Area Number	10178104
Research Institution	Osaka University
Principal Investigator	谷口直之大阪大学, 医学系研究科, 教授 (90002188)
Co-Investigator(Kenkyū-buntansha)	鈴木明身理化学研究所, 国際フロンティア, グループリーダー (70134533) 神奈木玲児大阪大学, 愛知県癌センター, 部長 (80161389) 木下タロウ大阪大学, 微生物研究所, 教授 (10153165) 辻崇一 (財)基礎腫瘍学研究会付属腫溜研究所, 研究員 (90124677) 古川鋼一名古屋大学, 医学部, 教授 (80211530)
Keywords	糖鎖 / 糖脂質 / 糖タンパク質 / 複合糖質 / 糖鎖リモデリング / 糖鎖遺伝子 / 糖転移酵素
Research Abstract	N-グリカンにbisectingGlcNAcを導入する糖転移酵素、GnT-IIIを導入した細胞ではEGFレセプターのエンドサイトーシスが亢進していること、またEGF刺激によるβ-カテニンのリン酸化が低下していることを見い出した。ドリコールリン酸マンノース合成酵素を制御するDPM2が、GPIアンカー合成の初期酵素をも制御することを見いだし、N型糖鎖とGPIアンカーの合成が連動していることを示した。細胞接着分子L-セレクチンの特異的リガンドと同定したシアリル6-スルホルイスxが、大腸上皮細胞に発現し、細胞の悪性化に伴って陰性化する傾向を有する事を見出した。これと対照的に、細胞が悪性化すると硫酸基を持たないシアリルルイスxの発現が増大する。正常大腸上皮細胞では、シアル酸の環状化によってセレクチンとの結合活性が調節されているが、悪性細胞ではこの調節機構が失われていることも判明した。ガングリオシド糖鎖合成に働く新しいシアル酸転移酵素遺伝子(ST6GalNAcV、VI)、およびグロボ系糖脂質合成に働くGb3合成酵素、Gb4合成酵素遺伝子を単離して機能を明らかにした。また、GM2/GD2合成酵素遺伝子ノックアウト、GD3合成酵素遺伝子ノックアウトマウスを用いて、複合型およびβ系列ガングリオシドが神経系維持と修復に必須なことを明らかにした。マウスシアル酸転移酵素のゲノム構造解析などのデータをまとめた。第3の新規Gal:→3硫酸転移酵素のcDNAクローニングを行い、O-結合型糖鎖のcore1鎖(Galβ1→3GalNAcα1→)に特異的に作用することを明らかにした。さらに基質特異性が既知のものとは異なる第4の酵素であるGal:→3硫酸転移酵素をブタ大腸から精製し、酵素学的性状を明らかにした。糖脂質硫酸転移酵素遺伝子のノックアウトマウスは、神経機能障害と精子形成障害をきたしたことから、硫酸化糖脂質のスルファチドとセミノリピドは、それぞれミエリン機能と精母細胞の分化に必須の分子であることが明らかとなった。

Research Products

(20 results)

All Other

All Publications (20 results)

[Publications] Tsuda T., et al.: "The Asn-420-linked sugar chain in human epidermal growth factor receptor suppresses ligand-independent spontaneous oligomerization : Possible role of a specific sugar chain in controllable receptor activation."J.Biol.Chem.. 275. 21988-21994 (2000)
[Publications] Taguchi T., et al.: "Purification and characterization of UDP-GlcNAc : GlcNAcβ1-6 (GlcNAcβ1-2) Man α 1-R [GlcNAc to Man] β1,4N-acetylglucosaminyltransferase VI from hen oviduct."J.Biol.Chem.. 275. 32598-32602 (2000)
[Publications] Sakamoto Y., et al.: "Molecular cloning and expression of cDNA encoding chicken UDP-GlcNAc : GlcNAcβ1-6 (GlcNAcβ1-2) Man α 1-R [GlcNAc to Man] β1,4N-acetylglucosaminyltransferase VI."J.Biol.Chem.. 275. 36029-36034 (2000)
[Publications] Sasai K., et al.: "The critical role of the stem region as a functional domain responsible for the oligomerization and Golgi localization of N-acetylglucosaminyltransferase V : The involvment of a domain homophilic interaction."J.Biol.Chem.. 276. 759-765 (2001)
[Publications] Kitada T., et al.: "The Addition of bisecting N-acetylglucosamine residues to E-cadherin downregulates the tyrosine phosphorylation of beta-catenin."J.Biol.Chem.. 276. 475-480 (2001)
[Publications] Honke K., et al.: "Molecular Cloning And Characterization of A Human β-Gal 3'-Sulfotransferase Which Acts on Both Type 1 and Type 2 (Gal β,3/1,4GlcNAc-R) Oligosaccharides."J.Biol.Chem.. 276. 267-274 (2001)
[Publications] Sato Y., et al.: "Overexpression of N-acetylglucosaminyltransferase III enhances the EGF-induced phosphorylation of ERK in HeLaS3 cells by upregulation of the internalization rate of the receptors."J.Biol.Chem.. (in press).
[Publications] Hong,Y. et al.: "Requirement of PIG-F and PIG-O for transferring phosphoethanolamine to the third mannose in glycosylphosphatidylinositol."J.Biol.Chem.. 275. 20911-20919 (2000)
[Publications] Watanabe,R. et al.: "Initial enzyme forglycosylphosphatidylinositol biosynthesis requires PIG-P and is regulated by DPM2."EMBO J.. 19. 4402-4411 (2000)
[Publications] Maeda,Y. et al.: "PIG-M transfers the first mannose to glycosylphosphatidylinositol on the lumenal side of the ER."EMBO J.. 20. 250-261 (2001)
[Publications] Kumamoto,K. et al.: "Increased expression of UDP-galactose transporter mRNA in human colon cancer tissues and its implication in synthesis of Thomsen-Friedenreich antigen and sialyl Lewis A/X determinants."Cancer Res.. (in press).
[Publications] Izawa,M., et al.: "Expression of sialyl 6-sulfo Lewis x is inversely correlated with conventional sialyl Lewis x expression in human colorectal cancer."Cancer Res.. 60. 1410-1416 (2000)
[Publications] Fukumoto,S. et al.: "GD3 synthase gene expression in PC12 cells results in the continuous activation of TrkA and ERK1/2 and enhanced proliferation."J.Biol.Chem.. 275. 5832-5838 (2000)
[Publications] Okajima,T. et al.: "Molecular cloning and expression of mouse GD1a/GT1aa/GQ1ba synthase (ST6GalNAc VI) gene."J.Biol.Chem.. 275. 6717-6723 (2000)
[Publications] Kojima,Y. et al.: "Molecular cloning of Gb3/CD77 synthase, a glycosyltransferase that initiate the synthesis of globo-series glycosphingolipids."J.Biol.Chem.. 275. 15152-15156 (2000)
[Publications] Honda,K. et al.: "Degradation of human aurora2 protein kinase by the anaphase-promoting complex-ubiquitin-proteasome pathway."Oncogene. 19. 2812-2819 (2000)
[Publications] Furukawa,K. et al.: "Molecular basis for the p phenotype : Identification of distinct and multiple mutations in the a1,4-galactosyltransferase gene in Swedisn and Japanese individuals."J.Biol.Chem.. 275. 37752-37756 (2000)
[Publications] Okajima,T. et al.: "Expression cloning of human globoside synthase cDNAs : Identification of b3Gal-T3 as UDP-N-acetylgalactosamine : globotriaosylceramide b1,3-N-acetylgalactosa-minyltransferase."J.Biol.Chem.. 275. 40498-40503 (2000)
[Publications] Hitoshi S. et al.: "Dorsal root ganglia neuron-specific promoter activity of the rabbit b-galactoside a1,2-fucosyltransferase gene."J.Biol.Chem.. 274. 389-396 (1999)
[Publications] Lee Y-C. et al.: "Molecular cloning and functional expression of two members of mouse NeuAca2,3Galb1,3GalNAc GalNAc a2,6-sialyltransferase family, ST6GalNAc III and IV."J.Biol.Chem.. 274. 11958-11967 (1999)

2000 Fiscal Year Annual Research Report

糖鎖遺伝子による細胞リモデリング

Principal Investigator

谷口 直之 大阪大学, 医学系研究科, 教授 (90002188)

Research Products

[Publications] Tsuda T., et al.: "The Asn-420-linked sugar chain in human epidermal growth factor receptor suppresses ligand-independent spontaneous oligomerization : Possible role of a specific sugar chain in controllable receptor activation."J.Biol.Chem.. 275. 21988-21994 (2000)

[Publications] Taguchi T., et al.: "Purification and characterization of UDP-GlcNAc : GlcNAcβ1-6 (GlcNAcβ1-2) Man α 1-R [GlcNAc to Man] β1,4N-acetylglucosaminyltransferase VI from hen oviduct."J.Biol.Chem.. 275. 32598-32602 (2000)

[Publications] Sakamoto Y., et al.: "Molecular cloning and expression of cDNA encoding chicken UDP-GlcNAc : GlcNAcβ1-6 (GlcNAcβ1-2) Man α 1-R [GlcNAc to Man] β1,4N-acetylglucosaminyltransferase VI."J.Biol.Chem.. 275. 36029-36034 (2000)

[Publications] Sasai K., et al.: "The critical role of the stem region as a functional domain responsible for the oligomerization and Golgi localization of N-acetylglucosaminyltransferase V : The involvment of a domain homophilic interaction."J.Biol.Chem.. 276. 759-765 (2001)

[Publications] Kitada T., et al.: "The Addition of bisecting N-acetylglucosamine residues to E-cadherin downregulates the tyrosine phosphorylation of beta-catenin."J.Biol.Chem.. 276. 475-480 (2001)

[Publications] Honke K., et al.: "Molecular Cloning And Characterization of A Human β-Gal 3'-Sulfotransferase Which Acts on Both Type 1 and Type 2 (Gal β,3/1,4GlcNAc-R) Oligosaccharides."J.Biol.Chem.. 276. 267-274 (2001)

[Publications] Sato Y., et al.: "Overexpression of N-acetylglucosaminyltransferase III enhances the EGF-induced phosphorylation of ERK in HeLaS3 cells by upregulation of the internalization rate of the receptors."J.Biol.Chem.. (in press).

[Publications] Hong,Y. et al.: "Requirement of PIG-F and PIG-O for transferring phosphoethanolamine to the third mannose in glycosylphosphatidylinositol."J.Biol.Chem.. 275. 20911-20919 (2000)

[Publications] Watanabe,R. et al.: "Initial enzyme forglycosylphosphatidylinositol biosynthesis requires PIG-P and is regulated by DPM2."EMBO J.. 19. 4402-4411 (2000)

[Publications] Maeda,Y. et al.: "PIG-M transfers the first mannose to glycosylphosphatidylinositol on the lumenal side of the ER."EMBO J.. 20. 250-261 (2001)

[Publications] Kumamoto,K. et al.: "Increased expression of UDP-galactose transporter mRNA in human colon cancer tissues and its implication in synthesis of Thomsen-Friedenreich antigen and sialyl Lewis A/X determinants."Cancer Res.. (in press).

[Publications] Izawa,M., et al.: "Expression of sialyl 6-sulfo Lewis x is inversely correlated with conventional sialyl Lewis x expression in human colorectal cancer."Cancer Res.. 60. 1410-1416 (2000)

[Publications] Fukumoto,S. et al.: "GD3 synthase gene expression in PC12 cells results in the continuous activation of TrkA and ERK1/2 and enhanced proliferation."J.Biol.Chem.. 275. 5832-5838 (2000)

[Publications] Okajima,T. et al.: "Molecular cloning and expression of mouse GD1a/GT1aa/GQ1ba synthase (ST6GalNAc VI) gene."J.Biol.Chem.. 275. 6717-6723 (2000)

[Publications] Kojima,Y. et al.: "Molecular cloning of Gb3/CD77 synthase, a glycosyltransferase that initiate the synthesis of globo-series glycosphingolipids."J.Biol.Chem.. 275. 15152-15156 (2000)

[Publications] Honda,K. et al.: "Degradation of human aurora2 protein kinase by the anaphase-promoting complex-ubiquitin-proteasome pathway."Oncogene. 19. 2812-2819 (2000)

[Publications] Furukawa,K. et al.: "Molecular basis for the p phenotype : Identification of distinct and multiple mutations in the a1,4-galactosyltransferase gene in Swedisn and Japanese individuals."J.Biol.Chem.. 275. 37752-37756 (2000)

[Publications] Okajima,T. et al.: "Expression cloning of human globoside synthase cDNAs : Identification of b3Gal-T3 as UDP-N-acetylgalactosamine : globotriaosylceramide b1,3-N-acetylgalactosa-minyltransferase."J.Biol.Chem.. 275. 40498-40503 (2000)

[Publications] Hitoshi S. et al.: "Dorsal root ganglia neuron-specific promoter activity of the rabbit b-galactoside a1,2-fucosyltransferase gene."J.Biol.Chem.. 274. 389-396 (1999)

[Publications] Lee Y-C. et al.: "Molecular cloning and functional expression of two members of mouse NeuAca2,3Galb1,3GalNAc GalNAc a2,6-sialyltransferase family, ST6GalNAc III and IV."J.Biol.Chem.. 274. 11958-11967 (1999)

谷口直之大阪大学, 医学系研究科, 教授 (90002188)