新規細胞内型ホスホリパーゼA1による逆行性輸送制御機構の解明

• Project/Area Number: 18050009
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Tohoku University (2007); The University of Tokyo (2006)
• Principal Investigator: 青木 淳賢 Tohoku University, 大学院・薬学研究科, 教授 (20250219)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥6,000,000 (Direct Cost: ¥6,000,000); Fiscal Year 2007: ¥3,000,000 (Direct Cost: ¥3,000,000); Fiscal Year 2006: ¥3,000,000 (Direct Cost: ¥3,000,000)
• Keywords: ホスホリパーゼA1 / 細胞内輸送 / 非対称分裂 / 線虫 / RNA干渉法 / ホスホリパーゼ

Research Abstract

ホスホリパーゼA_1(PLA_1)は、リン脂質のアシル鎖のうち1位を加水分解し2-アシルリゾリン脂質を産生する酵素である。その活性自体は古くから知られているが、PLA_1の生体内機能はほとんど不明である。細胞内型PLA_1(iPLA_1)は真核生物に保存されたファミリーを形成しており、線虫には1つの、哺乳類には3種類のiPLA_1(PA-PLA_1、p125、KIAA0725)が存在する。我々は、これらiPLA_1の生体内機能、さらには産物である2-アシルリゾリン脂質に焦点をあて研究を行っている。特に今回哺乳類iPLA_1,KIAA0725と線虫iPLA_1の機能解析を行った。
KIAA0725の機能解析
siRNAによるKIAA0725の発現抑制によりCTXのゴルジ体からERへの輸送速度が顕著に低下していることがわかった。また、強制発現細胞内には、2-アシルリゾホスファチジルエタノールアミン(1ysoPE)の蓄積が観察された。
一方、siRNAによるKIAA0725の発現抑制によりKIAA0725がオートファジーに関与する知見が得られた。
線虫iPLA_1の解析
まず、UV/TMP法を用い作製された、ipla-1の遺伝子欠損C.elegansを解析し、ipla-1欠損変異体は幹細胞様上皮細胞(seam cell)の非対称分裂のうち最後の細胞分裂のみに異常が生じるというユニークな表現型を示した。

Research Products

• [Journal Article] Protection against oxidative stress-induced hepatic injury by intracellular type II PAF-acetylhydrolase by metabolism of oxidized phospholipids in vivo. (2008)
  • Author(s): Kono N, Inoue T, Yoshida Y, Sato H, Matsusue T, Itabe H, Niki E, Aoki J, Arai H.
  • Journal Title: J. Biol. Chem. 283 Pages: 1628-1636
  • Peer Reviewed
• [Journal Article] Analysis of serum and urinary lysophospholipase D/autotaxin in nephrotic syndrome. (2008)
  • Author(s): Nakamura K, Nangaku M, Ohkawa R, Okubo S, Yokota H, Ikeda H, Aoki J, Yatomi Y.
  • Journal Title: Clin. Chem. Lab. Med. 46 Pages: 150-151
  • Peer Reviewed
• [Journal Article] Validation of an autotaxin enzyme immunoassay in human serum samples and its application to hypoalbuminemia differentiation. (2008)
  • Author(s): Nakamura K, Igarashi K, Ide K, Ohkawa R, Okubo S, Yokota H, Masuda A, Oshima N, Takeuchi T, Nangaku M, Okudaira S, Arai H, Ikeda H, Aoki J, Yatomi Y.
  • Journal Title: Clin, Chim. Acta 388 Pages: 51-58
  • Peer Reviewed
• [Journal Article] Embryo Spacing and Implantation Timing Are Differentially Regulated by LPA3-Mediated Lysophosphatidic Acid Signaling in Mice. (2007)
  • Author(s): Hama K, Aoki J, Inoue A, Endo T, Amano T, Motoki R, Kanai M, Ye X, Chun J, Matsuki N, Suzuki H, Shibasaki M, Arai H.
  • Journal Title: Biol. Reprod 77 Pages: 954-959
  • Peer Reviewed
• [Journal Article] Phospholipase A(1)assays using a radiolabeled substrate and mass spectrometry. (2007)
  • Author(s): Morikawa R, Tsujimoto M, Arai H, Aoki J.
  • Journal Title: Methods Enzymol. 434 Pages: 1-13
  • Peer Reviewed
• [Journal Article] Serum lysophospholipase D/autotaxin may be a new nutritional assessment marker: study on prostate cancer patients. (2007)
  • Author(s): Nakamura K, Takeuchi T, Ohkawa R, Okubo S, Yokota H, Tozuka M, Aoki J, Arai H, Ikeda H, Ohshima N, Kitamura T, Yatomi Y.
  • Journal Title: Ann. Clin. Biochem. 44 Pages: 549-556
  • Peer Reviewed
• [Journal Article] Type I platelet-activating factor acetylhydrolase catalytic subunits over-expression induces pleiomorphic nuclei and centrosome amplification. (2007)
  • Author(s): Yamaguchi N, Koizumi H, Aoki J, Natori Y, Nishikawa K, Natori Y, Takanezawa Y, Arai H.
  • Journal Title: Genes Cells 12 Pages: 1153-1161
  • Peer Reviewed
• [Journal Article] Plasma lysophosphatidic acid level and serum autotaxin activity are increased in liver injury in rats in relation to its severity. (2007)
  • Author(s): Watanabe N, Ikeda H, Nakamura K, Ohkawa R, Kume Y, Tomiya T, Tejima K, Nishikawa T, Arai M, Yanase M, Aoki J, Arai H, Omata M, Fujiwara K, Yatomi Y.
  • Journal Title: Life Sci 81 Pages: 1009-1015
  • Peer Reviewed
• [Journal Article] Submucosal connective tissue-type mast cells contribute to the production of lysophosphatidic acid (LPA) in the gastrointestinal tract through the secretion of autotaxin (ATX) /lysophospholipase D (lysoPLD). (2007)
  • Author(s): Mori K, Kitayama J, Aoki J, Kishi Y, Shida D, Yamashita H, Arai H, Nagawa H.
  • Journal Title: Virchows Arch. 451 Pages: 47-56
  • Peer Reviewed
• [Journal Article] Lysophosphatidic acid protection against apoptosis in the human pre-B-cell line Nalm-6. (2007)
  • Author(s): Satoh Y, Ohkawa R, Nakamura K, Higashi K, Kaneko M, Yokota H, Aoki J, Arai H, Yuasa Y, Yatomi Y.
  • Journal Title: Eur. J. Haematol. 78 Pages: 510-517
  • Peer Reviewed
• [Journal Article] Diversified expression patterns of autotaxin, a gene for phospholipid-generating enzymeduring mouse and chicken development. (2007)
  • Author(s): Ohuchi H, Hayashibara Y, Matsuda H, Onoi M, Mitsumori M, Tanaka M, Aoki J, Arai H, Noji S.
  • Journal Title: Dev. Dyn. 236 Pages: 1134-1143
  • Peer Reviewed
• [Journal Article] Peritoneal fluids from patients with certain gynecologic tumor contain elevated levels of bioactive lysophospholipase D activity. (2007)
  • Author(s): Tokumura A, Kume T, Fukuzawa K, Tahara M, Tasaka K, Aoki J, Arai H
  • Journal Title: Life Sci. 80 Pages: 1641-1649
  • Peer Reviewed
• [Journal Article] Neuroprotective Role of Transgenic PAF-Acetylhydrolase II in Mouse Modele of Focal Cerebral Ischemia. (2007)
  • Author(s): Umemura K, Kato I, Hirashima Y, Ishii Y, Inoue T, Aoki J, Kono N, Oya T, Hayashi N, Hamada H, Endo S, Oda M, Arai H, Kinouchi H, Hiraga K.
  • Journal Title: Stroke 38 Pages: 1036-1068
  • Peer Reviewed
• [Journal Article] Plasma lysophosphatidic acid is increased in Iiver fibrosis in rats and human with enhanced serum autotaxin level. (2007)
  • Author(s): Watanabe N, Ikeda H, Nakamura K, Ohkawa R, Kume Y, AokiJ, Hama K, Okudaira S, Tanaka M, Yanase M, Nagashima K, Tomiya T, Arai M, Inoue Y, Tejima K, Nishikawa T, Arai H, Omata M, Fujiwara K, Yatomi Y
  • Journal Title: J. Clin. Gastroenterol 41 Pages: 616-623
  • Peer Reviewed
• [Journal Article] Structure and function of extracellular phospholipase Al belonging to the pancreatic lipase gene family (review). (2007)
  • Author(s): Aoki J, Inoue A, Makide K, Saiki N, Arai H.
  • Journal Title: Biochimie 89 Pages: 197-204
  • Peer Reviewed
• [Journal Article] Measurement of lysophospholipase D/autotaxin activity in human serum samples (2007)
  • Author(s): Nakamura K, Ohkawa R, Okubo S, Tozuka M, Okada M, Aoki S, Aoki J, Arai H, Ikeda H, Yatomi Y
  • Journal Title: Clin. Biochem. 40 Pages: 274-277
  • Peer Reviewed
• [Journal Article] Autotaxin (NPP-2) in the brain: Cell type-specific expression and regulation during development and after neurotrauma (2007)
  • Author(s): Savaskan NE, Rocha L, Kotter MR, Lubec G, Baer A, van Meeteren LA, Kishi Y, Aoki J, Moolenaar WH, Nitsch R, Brauer AU.
  • Journal Title: Cell. Mol. Life Sci. 64 Pages: 230-243
  • Peer Reviewed
• [Journal Article] Autotaxin stabilizes blood vessels and is required for embryonic vasculature by producing lysophosphatidic acid (2006)
  • Author(s): H.Sakagami et al.
  • Journal Title: J. Biol. Chem. 281 Pages: 25822-25830
• [Journal Article] Autotaxin is overexpressed in glioblastoma multiforme and contributes to cell motility of glioblastoma by converting lysophosphatidylcoholine to lysophosphatidic acid (2006)
  • Author(s): Y.Kishi et al.
  • Journal Title: J. Biol. Chem. 281 Pages: 17492-17500
• [Journal Article] Cyclic phosphatidic acid is produced by autotaxin in blood (2006)
  • Author(s): S.Tsuda et al.
  • Journal Title: J. Biol. Chem. 281 Pages: 26081-26088
• [Journal Article] Phospholipase A1 -structure, distribution and function- (2006)
  • Author(s): A.Inoue at al.
  • Journal Title: Future Lipidology 1 Pages: 687-700
• [Journal Article] Lysophosphatidic receptor, LPA3, is positively and negatively regulated by progesterone and estrogen in the mouse uterus (2006)
  • Author(s): K.Hama et al.
  • Journal Title: Life Sciences 79 Pages: 736-1740
• [Journal Article] Transgenic Expression of Group V, but not Group X, Secreted PLA2 in Mice Leads to Neonatal Lethality due to Lung Dysfunction (2006)
  • Author(s): M.Ohtsuki et al.
  • Journal Title: J. Biol. Chem. 281 Pages: 36420-36433