1998 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, 医学部, 教授 (90002188)
Co-Investigator(Kenkyū-buntansha)	辻崇一理化学研究所, 国際フロンティア, チームリーダー (90124677) 古川鋼一名古屋大学, 医学部, 教授 (80211530) 鈴木明身東京都臨床医学総合研究所, 部長 (70134533) 神奈木玲児愛知県がんセンター, 部長 (80161389) 木下タロウ大阪大学, 微生物病研究所, 教授 (10153165)
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	白血球系細胞においては、シアリルルイスxがセレクチンの主要なリガンドであり、これがフコシルトランスフェラーゼVIIによって発現が調節されること、また細胞の悪性化や刺激に際して、この酵素遺伝子の転写が活性化されることを明らかにした。一方、固形癌細胞、特に消化門癌では、シアリルルイスaがシアリルルイスxと並んで重要なセレクチンのリガンドとして機能し、その発現が癌の血行性転移および患者の予後と深く関連することを明らかにした。リンパ節へのリンパ球のホーミングを媒介するL-セレクチンリガンドが、シアリル6-スルホルイスxであることを明らかにした。GM2/GD2合成酵素遺伝子、GD3合成酵素遺伝子のノックアウトマウスを作成し、その神経系、免疫系、生殖系の異常につき解析し報告した。GPIアンカーの生合成とタンパク質への転移に関与する遺伝子のうち、ドリコールリン酸マンノースを合成する酵素の遺伝子であるDPM1と、DPM2はそれぞれ触媒サブユニットと制御サブユニットをコードすることを証明した。Neuro2a細胞に誘導可能な形で導入したGD3合成酵素cDNAの発現により新たに発現してくる幾つかの遺伝子をDifferential display法により検索した結果、DAP-群であることが明らかになった。神経系や初期胚でLexを合成する酵素は、過去にクローンされたFUT3-7のいずれでもなく、新たにクローン化したFuc-TIXであることがわかった。Fuc-TIXの配列は種を越えてきわめてよく保存されており、発生的に重要な機能をはたしていることが予想された。NeuGcの発現を律速支配している酵素CMP-NeuAc水酸化酵素を精製し、クローニングした。ヒトではNeuGcは発現していないが、その原因が本酵素遺伝子の92bp配列の欠失であることをを明らかにした。遺伝子座はマウスで13A3、ヒトで6p22に存在すること、赤毛ザルはヒトで見られる欠失は起きていないことを確認した。Nアセチルグルコサミン転移酵素トランスジェニックマウスを作製し、解析した結果、この酵素の異所性発現によりよりリポ蛋白質の分泌が障害を受け、肝に脂質が蓄積することを見い出した。

Research Products

(24 results)

All Other

All Publications (24 results)

[Publications] R.Watanabe et al.: "The first step of glycosylphosphatidylinositol biosynthesis is mediated by a complex of PIG-A,PIG-H,PIG-C and GPII." EMBOJ.17. 877-885 (1998)
[Publications] S.Tomita et al.: "A homologue of Saccharomyces cerevisiae Dpmlp is not sufficient for synthesis of dolichol-phosphate-mannose in mammalian cells." J.Biol.Chem.273. 9249-9254 (1998)
[Publications] Y.Maeda et al.: "DPM2 regulates biosynthesis of dolichol phosphate-mannose in mammalian cells : correct subcellular localization and stabilization of DPMI,and binding of dolichol phosphate." EMBOJ.17. 4920-4929 (1998)
[Publications] A.Irie et al.: "The molecular basis for the absence of N-glycolylneuraminic acid in humans." J.Biol.Chem.273. 15866-15871 (1998)
[Publications] S.Hitoshi et al.: "Dorsal root ganglia neuron-specific promoter activity of the rabbit β-galactoside α1,2-fucosyltransferase gene." J.Biol.Chem.274. 389-396 (1999)
[Publications] S.Takashima et al.: "Genomic structure and promoter activity of the mouse polysialic acid synthase(mST8Sia IV/PST)gene." J.Biol.Chem.273. 7675-7683 (1998)
[Publications] T.Okajima et al.: "Molecular cloning of a novel α2,3-sialyltransferase(ST3Gal VI)that sialylates type II lactosamine structures on glycoproteins and glycolipids." J.Biol.Chem.(in press).
[Publications] K.Honke,et al.: "Cancer-associated expression of glycolipid sulfotransferase gene in human renal cell carcinoma cells." Cancer Res.58. 3800-3805 (1998)
[Publications] K.Fukushima et al.: "Elevation of alfa2-6 sialyltransferase and alfal-2 fucosyltransferase activities in human choriocarcinoma." Cancer Res.58. 4301-4306 (1998)
[Publications] Y.Ihara et al.: "Ectopic expression of N-acetylglucosaminyltransferase III in transgenic hepatocytes disrupts apolipoprotein B secretion and induces aberrant cellular morphology with lipid atorage." Proc.Natl.Acad.Sci.USA.95. 2526-2530 (1998)
[Publications] Y.Sakamoto et al.: "Purification and characterization of UDP-GlcNAc : Galβ1-4GlcNAcβ1-3Galβ1-4Glc(NAc)-R(GlcNAc to Gal)β1,6N-acetylglucosaminyltransferase from hog small intestine." J.Biol.Chem.273. 27625-27632 (1998)
[Publications] E.Miyoshi et al.: "Overexpression of α1-6fucosyltransferase in hepatoma cells suppressed intrahepatic metastasis after splenic injection in athymic mice." Cancer Res.(in press).
[Publications] N.Kimura et al.: "Reconstitution of funcilonal L-selectin ligands on a cultured human endothelial cell line by co-transfection of α1->3 fucosyltransferase VII and newly cloned GlcNAcβ : 6-sulfotransferase cDNA." Proc.Natl.Acad.Sci.USA. (in press).
[Publications] M.Nakamura et al.: "Single glycosyltransferase,core 2 β-1,6-N-acetylglucosaminyltransferase,regulates cell surface sialyl-Lex expression level in human pre-B lymphocytle leukemia cell Hne KM3 treated with phorbolester." J.Biol.Chem.273. 26779-26789 (1998)
[Publications] T.Kudo et al.: "Expression cloning and characterization of a novel murine α1,3-fucosyltransferase,mFuc-TIX,that synthesizes the Lewis x(CD15)epitope in brain and kindney." J.Biol.Chem.273. 26729-26738 (1998)
[Publications] H.Narimatsu et al.: "Lewis and Secretor gene dosages affect CA19-9 and DU-PAN-2 serum levels in normal individuals and colorectal cancer patients." Cancer Res.58. 512-518 (1998)
[Publications] K.Takamiya et al.: "Complex gangliosides are essential in spermatogenesis of mice : Possible roles in transport of testosterone." Proc.Natl.Acad.Sci.USA. 95. 12147-12152 (1998)
[Publications] S.Fukumoto et al.: "Expression cloning of mouse cDNA of CMP-NeuAc : lactosylceramide α2,3-sialyltransferase(GM3 synthase),an enzyme that initiates the synthesis of gangliosides." J.Biol.Chem.(in press).
[Publications] M.Tanemura et al.: "Reduction of the major swine xenoantigen,the α-galactosyl epitope by transfection of the α2,3-sialyltransferase gene." J.Biol.Chem.273. 16421-16425 (1998)
[Publications] J.Mitoma et al.: "Occurrence of an unusual lipid,phosphatidyl-L-threone,in cultured hippocampal neurons : exogenous L-serine is requiredfor the synthesis of neuronal phosphatidyl-L-serine and sphingolipids" J.Biol.Chem.273. 19363-19366 (1998)
[Publications] H.Komori et al.: "Regulation of intracellular ceramide content in B16 melanoma cells : Biological implications of ceramide glycosylation" J.Biol.Chem.(in press).
[Publications] C.Mitsuoka et al.: "Identification of a major carbohydrate capping group of the L-selectin ligand on high endothelial venules in human lymph nodes as 6-sulfo sialyl Lewis x." J.Biol.Chem.273. 11225-11233 (1998)
[Publications] K.Uchimura et al.: "Molecular cloning and characterization of an N-acetylglucosamine-6-O-sulfotransferase." J.Biol.Chem.273. 22577-22583 (1998)
[Publications] C.Mitsuoka et al.: "Regulation of selectin binding activity by cyclization of sialic acid moiety of carbohydrate ligands on human leukocytes." Proc.Natl.Acad.Sci.USA. 96. 1597-1602 (1999)

1998 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, 医学部, 教授 (90002188)

Research Products

[Publications] R.Watanabe et al.: "The first step of glycosylphosphatidylinositol biosynthesis is mediated by a complex of PIG-A,PIG-H,PIG-C and GPII." EMBOJ.17. 877-885 (1998)

[Publications] S.Tomita et al.: "A homologue of Saccharomyces cerevisiae Dpmlp is not sufficient for synthesis of dolichol-phosphate-mannose in mammalian cells." J.Biol.Chem.273. 9249-9254 (1998)

[Publications] Y.Maeda et al.: "DPM2 regulates biosynthesis of dolichol phosphate-mannose in mammalian cells : correct subcellular localization and stabilization of DPMI,and binding of dolichol phosphate." EMBOJ.17. 4920-4929 (1998)

[Publications] A.Irie et al.: "The molecular basis for the absence of N-glycolylneuraminic acid in humans." J.Biol.Chem.273. 15866-15871 (1998)

[Publications] S.Hitoshi et al.: "Dorsal root ganglia neuron-specific promoter activity of the rabbit β-galactoside α1,2-fucosyltransferase gene." J.Biol.Chem.274. 389-396 (1999)

[Publications] S.Takashima et al.: "Genomic structure and promoter activity of the mouse polysialic acid synthase(mST8Sia IV/PST)gene." J.Biol.Chem.273. 7675-7683 (1998)

[Publications] T.Okajima et al.: "Molecular cloning of a novel α2,3-sialyltransferase(ST3Gal VI)that sialylates type II lactosamine structures on glycoproteins and glycolipids." J.Biol.Chem.(in press).

[Publications] K.Honke,et al.: "Cancer-associated expression of glycolipid sulfotransferase gene in human renal cell carcinoma cells." Cancer Res.58. 3800-3805 (1998)

[Publications] K.Fukushima et al.: "Elevation of alfa2-6 sialyltransferase and alfal-2 fucosyltransferase activities in human choriocarcinoma." Cancer Res.58. 4301-4306 (1998)

[Publications] Y.Ihara et al.: "Ectopic expression of N-acetylglucosaminyltransferase III in transgenic hepatocytes disrupts apolipoprotein B secretion and induces aberrant cellular morphology with lipid atorage." Proc.Natl.Acad.Sci.USA.95. 2526-2530 (1998)

[Publications] Y.Sakamoto et al.: "Purification and characterization of UDP-GlcNAc : Galβ1-4GlcNAcβ1-3Galβ1-4Glc(NAc)-R(GlcNAc to Gal)β1,6N-acetylglucosaminyltransferase from hog small intestine." J.Biol.Chem.273. 27625-27632 (1998)

[Publications] E.Miyoshi et al.: "Overexpression of α1-6fucosyltransferase in hepatoma cells suppressed intrahepatic metastasis after splenic injection in athymic mice." Cancer Res.(in press).

[Publications] N.Kimura et al.: "Reconstitution of funcilonal L-selectin ligands on a cultured human endothelial cell line by co-transfection of α1->3 fucosyltransferase VII and newly cloned GlcNAcβ : 6-sulfotransferase cDNA." Proc.Natl.Acad.Sci.USA. (in press).

[Publications] M.Nakamura et al.: "Single glycosyltransferase,core 2 β-1,6-N-acetylglucosaminyltransferase,regulates cell surface sialyl-Lex expression level in human pre-B lymphocytle leukemia cell Hne KM3 treated with phorbolester." J.Biol.Chem.273. 26779-26789 (1998)

[Publications] T.Kudo et al.: "Expression cloning and characterization of a novel murine α1,3-fucosyltransferase,mFuc-TIX,that synthesizes the Lewis x(CD15)epitope in brain and kindney." J.Biol.Chem.273. 26729-26738 (1998)

[Publications] H.Narimatsu et al.: "Lewis and Secretor gene dosages affect CA19-9 and DU-PAN-2 serum levels in normal individuals and colorectal cancer patients." Cancer Res.58. 512-518 (1998)

[Publications] K.Takamiya et al.: "Complex gangliosides are essential in spermatogenesis of mice : Possible roles in transport of testosterone." Proc.Natl.Acad.Sci.USA. 95. 12147-12152 (1998)

[Publications] S.Fukumoto et al.: "Expression cloning of mouse cDNA of CMP-NeuAc : lactosylceramide α2,3-sialyltransferase(GM3 synthase),an enzyme that initiates the synthesis of gangliosides." J.Biol.Chem.(in press).

[Publications] M.Tanemura et al.: "Reduction of the major swine xenoantigen,the α-galactosyl epitope by transfection of the α2,3-sialyltransferase gene." J.Biol.Chem.273. 16421-16425 (1998)

[Publications] J.Mitoma et al.: "Occurrence of an unusual lipid,phosphatidyl-L-threone,in cultured hippocampal neurons : exogenous L-serine is requiredfor the synthesis of neuronal phosphatidyl-L-serine and sphingolipids" J.Biol.Chem.273. 19363-19366 (1998)

[Publications] H.Komori et al.: "Regulation of intracellular ceramide content in B16 melanoma cells : Biological implications of ceramide glycosylation" J.Biol.Chem.(in press).

[Publications] C.Mitsuoka et al.: "Identification of a major carbohydrate capping group of the L-selectin ligand on high endothelial venules in human lymph nodes as 6-sulfo sialyl Lewis x." J.Biol.Chem.273. 11225-11233 (1998)

[Publications] K.Uchimura et al.: "Molecular cloning and characterization of an N-acetylglucosamine-6-O-sulfotransferase." J.Biol.Chem.273. 22577-22583 (1998)

[Publications] C.Mitsuoka et al.: "Regulation of selectin binding activity by cyclization of sialic acid moiety of carbohydrate ligands on human leukocytes." Proc.Natl.Acad.Sci.USA. 96. 1597-1602 (1999)

谷口直之大阪大学, 医学部, 教授 (90002188)