Role of BIG2, a Guanine-Nucleotide Exchanging factor for ADP-Ribosylation Factors, in Insulin-Regulated Glucose Transporter Translocation

• Project/Area Number: 21570142
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: The University of Tokushima
• Principal Investigator: UCHIYAMA Keiji 徳島大学, 疾患酵素学研究センター, 准教授 (60294039)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2011: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2010: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2009: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: Glut4 / BIG2 / Glut4トランスロケーション / インスリン作用 / 脂肪細胞 / インスリン / Glut4小胞再構成 / リサイクリングエンドソーム / 3T3-L1脂肪

Research Abstract

We here show that BIG2 plays an important role in a formation of Glut4 vesicles from endosomal compartment in both basal and insulin-stimulated adipocytes. In 3T3-L1 adipocytes, knockdown of BIG2 by RNA interference leaded to insufficient insulin-stimulated Glut4 translocation to the plasma membrane. The basal distribution of Glut4 was analyzed by subcellular fractionation. It was shown that the formation of Glut4-vesicles in the BIG2-knockdown cells was strongly repressed. Here, we used an in vitro reconstitution of Glut4-vesicles in order to demonstrate that these Glut4-vesicles are formed from endosomal membranes in a BIG2-dependent manner. Reducing endogenous BIG2 inhibited in vitro formation of Glut4 vesicles from the endosomal compartment prepared from not only unstimulated but insulin-stimulated adipocytes.

Research Products

• [Journal Article] VCIP135 deubiquitinase and its binding protein, WAC, in p97ATPase-mediated membrane fusion (2011)
  • Author(s): Totsukawa G, Kaneko Y, Uchiyama K, Toh H, Tamura K, and Kondo H
  • Journal Title: The EMBO Journal Volume: Vol.30, No.17 Pages: 3455-3474
  • Peer Reviewed
• [Journal Article] VCIP135 deubiquitinase and its binding proteinon (2011)
  • Author(s): Totsukawa G, Kaneko Y, Uchiyama K, TohH, Kondo H
  • Journal Title: WAC Volume: 30(17) Issue: 17 Pages: 3581-3593
  • DOI: 10.1038/emboj.2011.260
  • Peer Reviewed
• [Journal Article] The Rab GTPase-Activating Protein AS160 as A Common Regulator of Insulin-and G. q-Mediated Intracellular GLUT4 Vesicle Distribution (2009)
  • Author(s): Yuasa, T, Uchiyama K., Ogura, Y., Kimura M., Teshigawara K., Hosaka T., Tanaka, Y., Obata T., Sano H., Kishi K., and Ebina Y.
  • Journal Title: Endocrine J. Volume: Vol.56, No.3 Pages: 345-359
  • Peer Reviewed
• [Journal Article] The Rab GTPase-Activating Protein AS160 as A Common Regulator of Insulin- and Gαq-Mediated Intracellular GLUT4 Vesicle DistributionA (2009)
  • Author(s): Yuasa T.
  • Journal Title: Endocrine Journal 56(3) Pages: 345-359
  • Peer Reviewed
• [Presentation] Functional roles of TUG and p97/ VCP in Glut4 trafficking (2012)
  • Author(s): Uchiyama, K. and Ebina, Y.
  • Organizer: 第34回分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜市)
  • Year and Date: 2012-12-16
• [Presentation] Functional roles of TUG and p97/VCP in Glut4 trafficking (2011)
  • Author(s): Keiji Uchiyama
  • Organizer: 第34回分子生物学会年会
  • Place of Presentation: パシフィコ横浜(神奈川県)(口頭発表)
  • Year and Date: 2011-12-16
• [Presentation] Functional roles of TUG and p97/VCP in Glut4 trafficking (2011)
  • Author(s): Keiji Uchiyama
  • Organizer: 第34回分子生物学会年会
  • Place of Presentation: パシフィコ横浜(神奈川県)(ポスター発表)
  • Year and Date: 2011-12-15
• [Presentation] プリオン感染細胞における細胞内小胞輸送の抑制 (2011)
  • Author(s): 内山圭司, 坂口末廣
  • Organizer: 第26回中国四国ウイルス研究会
  • Place of Presentation: 徳島大学(徳島市)
  • Year and Date: 2011-06-20
• [Presentation] プリオン感染細胞における細胞内小胞輸送の抑制 (2011)
  • Author(s): 内山圭司
  • Organizer: 第26回中国四国ウイルス研究会
  • Place of Presentation: 徳島大学(徳島県)
  • Year and Date: 2011-06-20
• [Presentation] Role of BIG2, a guanine-nucleotide exchanging factor for ADP-Ribosylation favtors, in insulin-regulated glucose transporter translocation (2009)
  • Author(s): Uchiyama, K. and Ebina, Y.
  • Organizer: 第32回分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜市)
  • Year and Date: 2009-12-09
• [Presentation] Role of BIG2, a guanine-nucleotide exchanging factor for ADP-Ribosylation favtors, in insulin-regulated glucose transporter translocation. (2009)
  • Author(s): 内山圭司
  • Organizer: 分子生物学会年会
  • Place of Presentation: 横浜(パシフィコ横浜)
  • Year and Date: 2009-12-09
• [Remarks]
  • URL: http://www.tokushima-u.ac.jp/ier/
• [Remarks]
  • URL: http://www.tokushima-u.ac.jp/ier/