Spatio-temporal analysis of DOCK180 during dorsal ruffle formation

• Project/Area Number: 24770127
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Functional biochemistry
• Research Institution: Kyushu University
• Principal Investigator: SANEMATSU FUMIYUKI 九州大学, 生体防御医学研究所, 助教 (80381094)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2013: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000); Fiscal Year 2012: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: ラッフル膜形成 / Rac活性化分子 / DOCK180 / 細胞骨格 / Rac / ホスファチジン酸 / ラッフル膜

Research Abstract

Activation of receptor tyrosine kinases leads to the formation of two different types of plasma membrane structures: peripheral ruffles and dorsal ruffles. Although the formation of both ruffle types requires activation of the small GTPase Rac, the difference in kinetics suggests that a distinct regulatory mechanism operates for their ruffle formation. DOCK1(DOCK180) and DOCK5 are atypical Rac activators and are both expressed in mouse embryonic fibroblasts (MEFs). We found that DOCK1 deficiency alone impaired dorsal ruffle formation in MEFs. DOCK1 bound to phosphatidic acid (PA) through the C-terminal polybasic amino acid cluster and was localized to dorsal ruffles. In addition, we show that phospholipase D, an enzyme that catalyzes PA synthesis, is required for PDGF-induced dorsal, but not peripheral, ruffle formation. These results indicate that the phospholipase D-PA axis selectively controls dorsal ruffle formation by regulating DOCK1 localization.

Research Products

• [Journal Article] The Rac activator DOCK2 regulates natural killer cell-mediated cytotoxicity in mice through the lytic synapse formation (2013)
  • Author(s): Sakai, Y., Tanaka, Y., Yanagihara, T., Watanabe, M., Duan, X., Terasawa, M., Nishikimi, A., Sanematsu, F., Fukui. Y.
  • Journal Title: Blood Volume: 122(3) Pages: 386-93
  • Peer Reviewed
• [Journal Article] Phosphatidic acid-dependent recruitment and function of the Rac activator DOCK1 during dorsal ruffle formation (2013)
  • Author(s): Sanematsu, F., Nishikimi, A., Watanabe, M., Hongu, T., Tanaka, Y., Kanaho, Y., Côté, J.F., Fukui, Y.
  • Journal Title: J Biol. Chem. Volume: 288(12) Pages: 8092-100
  • Peer Reviewed
• [Journal Article] The Rac activator DOCK2 regulates natural killer cell-mediated cytotoxicity in mice through the lytic synapse formation (2013)
  • Author(s): Sakai Y, Tanaka Y, Yanagihara T, Watanabe M, Duan X, Terasawa M, Nishikimi A, Sanematsu F, Fukui Y.
  • Journal Title: Blood Volume: 122 Issue: 3 Pages: 386-393
  • DOI: 10.1182/blood-2012-12-475897
  • Peer Reviewed
• [Journal Article] Phosphatidic acid-dependent recruitment and function of the Rac activator DOCK1 during dorsal ruffle formation. (2013)
  • Author(s): Sanematsu F, Nishikimi A, Watanabe M, Hongu T, Tanaka Y, Kanaho Y, Cote JF, Fukui Y
  • Journal Title: J. Biol. Chem. Volume: 288 Issue: 12 Pages: 8092-8100
  • DOI: 10.1074/jbc.m112.410423
  • Peer Reviewed
• [Journal Article] 心血管形成におけるDOCK180の機能とその制御機構 (2013)
  • Author(s): 實松史幸
  • Journal Title: 感染 炎症 免疫 Volume: 42 Pages: 50-53
• [Journal Article] Dimerization of DOCK2 is essential for DOCK2-mediated Rac activation and lymphocyte migration (2012)
  • Author(s): Terasawa, M., Uruno, T., Mori, S., Kukimoto-Niino, M., Nishikimi, A., Sanematsu, F., Tanaka, Y., Yokoyama, S., Fukui, Y.
  • Journal Title: PLoS One Volume: 7(9)
  • Peer Reviewed
• [Journal Article] Blockade of inflammatory responses by a small-molecule inhibitor of the Rac activator DOCK2 (2012)
  • Author(s): Nishikimi, A., Uruno, T., Duan, X., Cao, Q., Okamura, Y., Saitoh, T., Saito, N., Sakaoka. S., Du, Y., Suenaga, A., Kukimoto-Niino, M., Miyano, K., Gotoh, K., Okabe, T., Sanematsu, F., Tanaka, Y., Sumimoto, H., Honma, T., Yokoyama, S., Nagano, T., Kohda, D., Kanai, M., Fukui, Y.
  • Journal Title: Chem. Biol. Volume: 19(4) Pages: 488-97
  • Peer Reviewed
• [Journal Article] DOCK8 is a Cdc42 activator critical for interstitial dendritic cell migration during immune responses (2012)
  • Author(s): Harada, Y., Tanaka, Y., Terasawa, M., Pieczyk, M., Habiro, K., Katakai, T., Hanawa-Suetsugu, K., Kukimoto-Niino, M., Nishizaki, T., Shirouzu, M, Duan, X., Uruno, T., Nishikimi, A., Sanematsu, F., Yokoyama, S., Stein, J.V., Kinashi, T., Fukui, Y.
  • Journal Title: Blood Volume: 119(19) Pages: 4451-61
  • Peer Reviewed
• [Journal Article] Dimerization of DOCK2 is essential for DOCK2-mediated Rac activation and lymphocyte migration (2012)
  • Author(s): Terasawa M, Uruno T, Mori S, Kukimoto-Niino M, Nishikimi A, Sanematsu F, Tanaka Y, Yokoyama S, Fukui Y
  • Journal Title: PLoS ONE Volume: 7 Issue: 9 Pages: e46277-e46277
  • DOI: 10.1371/journal.pone.0046277
  • Peer Reviewed
• [Journal Article] Blockade of inflammatory responses by a small-molecule inhibitor of the Rac activatorDOCK2 (2012)
  • Author(s): Nishikimi A, Uruno T, Duan X, Cao Q, Okamura Y, Saitoh T, Saitoh N, Sakaoka S, Du Y, Suenaga A, Kukimoto-Niino M, Miyano K, Gotoh K, Okabe T, Sanematsu F, Tanaka Y, Sumimoto H., Honma T, Yokoyama S, Nagano T, Kohda D, Kanai M, and Fukui Y
  • Journal Title: Chem. Biol Volume: 19 Issue: 4 Pages: 488-497
  • DOI: 10.1016/j.chembiol.2012.03.008
  • Peer Reviewed
• [Journal Article] DOCK8 is a Cdc42 activator critical for interstitial dendritic cell migrataon during immune responses (2012)
  • Author(s): Yosuke Harada, Yoshihiko Tanaka, Masao Terasawa, Markus Pieczyk, Katsuyoshi Habiro, Tomoya Katakai, Kyoko Hanawa-Suetsugu, Mutsuko Kukimoto-Niino, Tomoko Nishizaki, Mikako Shirouzu, Xuefeng Duan, Takehito Uruno, Akihiko Nishikimi, Fumiyuki Sanematsu, Shigeyuki Yokoyama, Jens V.Stein, Tatsuo Kinashi, Yoshinori Fukui
  • Journal Title: Blood Volume: (In press) Issue: 19 Pages: 4451-4461
  • DOI: 10.1182/blood-2012-01-407098
  • Peer Reviewed
• [Presentation] Dorsal ruffle形成時におけるDOCK1の役割とその制御 (2012)
  • Author(s): 實松史幸, 錦見昭彦, 渡邉真裕紀, 本宮綱記, 田中芳彦, 金保安則, Jean-Francois Cote, 福井宣規
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡
  • Year and Date: 2012-12-12
• [Presentation] Role of DOCK1 and Its Regulation during Dorsal ruffle formation.
  • Author(s): Sanematsu F et al.
  • Organizer: The 35th Annual Meeting of the Molecular Biology Society of Japan
  • Place of Presentation: Marinemesse Fukuoka, Japan
• [Remarks]
  • URL: http://www.bioreg.kyushu-u.ac.jp/iden/