Molecular characterization of a HECT-type E3 ubiquitin ligase that is encoded in the WWP1 gene responsible for chicken muscular dystrophy.

• Project/Area Number: 24500472
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Neurophysiology and muscle physiology
• Research Institution: National Center of Neurology and Psychiatry
• Principal Investigator: IMAMURA Michihiro 独立行政法人国立精神・神経医療研究センター, 神経研究所 遺伝子疾患治療研究部, 室長 (80221787)
• Co-Investigator(Kenkyū-buntansha): TAKEDA Shin'ichi 独立行政法人国立精神・神経医療研究センター神経研究所, 遺伝子疾患治療研究部, 部長 (90171644)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 筋ジストロフィー / 実験動物 / 遺伝子疾患 / WWP1 E3 ユビキチンリガーゼ / タンパク質分解 / WWP1 / E3ユビキチンリガーゼ / ミスセンス変異 / 筋形質膜 / 筋小胞 / ミトコンドリア / HECT型E3ユビキチンリガーゼ / ジストログリカン / サルコグリカン / 骨格筋 / HECTドメイン / WWドメイン

Outline of Final Research Achievements

The R441Q missense mutation in the WWP1 protein was reported as the most promising candidate responsible for chicken muscular dystrophy (MD). We generated an antibody against WWP1, and characterized WWP1 protein expression in skeletal muscles in vivo and in vitro. In wild-type skeletal muscle, WWP1 was detected as an approximately 130-kDa protein and localized to the sarcolemma, sarcoplasmic reticulum, mitochondria, as well as in a part of the nucleus, whereas it was markedly degraded and absent from the sarcolemma in MD muscle. These changes were already observed in MD chickens in the pre-pathological stage. In vitro expression analysis demonstrated significant degradation of mutant but not wild-type WWP1, specifically in myogenic cells. Our study revealed that the R441Q missense mutation in the WWP1 protein causes its degradation as well as loss of its sarcolemmal localization, which are hence implicated in the pathogenesis of chicken MD.

Research Products

• [Journal Article] Low intensity training of mdx mice reduces carbonylation and increases expression levels of proteins involved in energy metabolism and muscle contraction. (2015)
  • Author(s): Hyzewicz J, Tanihata J, Kuraoka M, Ito N, Miyagoe-Suzuki Y, Takeda S
  • Journal Title: Free Radic Biol Med Volume: 82 Pages: 122-136
  • DOI: 10.1016/j.freeradbiomed.2015.01.023
  • Peer Reviewed / Open Access
• [Journal Article] Fukutin is prerequisite to ameliorate muscular dystrophic phenotype by myofiber-selective LARGE expression. (2015)
  • Author(s): Ohtsuka Y, Kanagawa M, Yu CC, Ito C, Chiyo T, Kobayashi K, Okada T, Takeda S’I, Toda T.
  • Journal Title: Sci Rep Volume: 5 Issue: 1 Pages: 8316-8316
  • DOI: 10.1038/srep08316
  • NAID: 120005600773
  • Peer Reviewed / Open Access
• [Journal Article] α1-Syntrophin deficient mice exhibit impaired muscle force recovery after osmotic shock. (2014)
  • Author(s): Yokota T, Miyagoe-Suzuki Y, Ikemoto T, Matsuda R, Takeda S
  • Journal Title: Muscle Nerve Volume: 49 Issue: 5 Pages: 728-735
  • DOI: 10.1002/mus.23990
  • Peer Reviewed
• [Journal Article] DOK7 gene therapy benefits mouse models of diseases characterized by defects in the neuromuscular junction (2014)
  • Author(s): Sumimasa Arimura, Takashi Okada, Tohru Tezuka, Tomoko Chiyo, Yuko Kasahara, Toshiro Yoshimura, Masakatsu Motomura, Nobuaki Yoshida, David Beeson, Shin’ichi Takeda, Yuji Yamanashi
  • Journal Title: Science Volume: 345 Issue: 6203 Pages: 1505-1508
  • DOI: 10.1126/science.1250744
  • Peer Reviewed / Open Access
• [Journal Article] Initial Pulmonary Respiration Causes Massive Diaphragm Damage and Hyper-CKemia in Duchenne Muscular Dystrophy Dog (2013)
  • Author(s): Nakamura A, Kobayashi M, Kuraoka M, Yuasa K, Yugeta N, Okada T, Takeda S
  • Journal Title: Sci Rep Volume: 3 Issue: 1 Pages: 2183-2183
  • DOI: 10.1038/srep02183
  • Peer Reviewed
• [Journal Article] Highly efficient in vivo delivery of PMO into regenerating myotubes and rescue in laminin-α2 chain-null congenital muscular dystrophy mice (2013)
  • Author(s): Aoki Y, Nagata T, Yokota T, Nakamura A, Wood MJ, Partridge T, Takeda S
  • Journal Title: Hum Mol Genet Volume: 22 Issue: 24 Pages: 4914-4928
  • DOI: 10.1093/hmg/ddt341
  • Peer Reviewed
• [Journal Article] Prednisolone improves walking in Japanese Duchenne muscular dystrophy patients (2013)
  • Author(s): Takeuchi F, Yonemoto N, Nakamura H, Shimizu R, Komaki H, Mori-Yoshimura M, Hayashi YK, Nishino I, Kawai M, Kimura E, Takeda S
  • Journal Title: J Neurol Volume: 260 Issue: 12 Pages: 3023-3029
  • DOI: 10.1007/s00415-013-7104-y
  • Peer Reviewed
• [Journal Article] Demonstration of systemic exon 45-55 multiple skipping in dystrophic mdx52 mice (2012)
  • Author(s): Aoki Y, Yokota T, Nagata T, Nakamura A, Nagaraju K, Hoffman E, Partridge T, Takeda S.
  • Journal Title: Proc Natl Acad Sci USA Volume: 109 Issue: 34 Pages: 13763-13768
  • DOI: 10.1073/pnas.1204638109
  • Peer Reviewed
• [Journal Article] Extensive and prolonged restoration of dystrophin expression with vivo-morpholino-mediated multiple exon skipping in dystrophic dogs. (2012)
  • Author(s): Yokota T, Nakamura A, Saito T, Nagata T, Partridge T, Hoffman E, Takeda S.
  • Journal Title: Nucleic Acid Ther Volume: 22 Issue: 5 Pages: 306-315
  • DOI: 10.1089/nat.2012.0368
  • Peer Reviewed
• [Presentation] Differences in Degradation and Distribution of Mutated WP1 E3 Ubiquitin Protein Ligase between Chicken and Mouse Skeletal Muscles (2014)
  • Author(s): Imamura, M., Takeda, S.
  • Organizer: American Society for Cell Biology
  • Place of Presentation: Pennsylvania Convention Center, Philadelphia, PA, U.S.A.
  • Year and Date: 2014-12-06 – 2014-12-10
• [Presentation] Analysis of Interaction of WWP1 E3 Ubiquitin Protein Ligase with Dystrophin-Associated Glycoproteins (2013)
  • Author(s): Imamura M, Takeda S
  • Organizer: 53rd American Society for Cell Biology Annual Meeting
  • Place of Presentation: Ernest N. Morial Convention Center, New Orleans, Louisiana, USA
• [Presentation] An R441Q mutation of the WWP1 gene causes WWP1 degradation in skeletal muscle of chicken muscular dystrophy (2012)
  • Author(s): Imamura M and Takeda S
  • Organizer: The American Society for Cell Biology 2012 Annual Meeting
  • Place of Presentation: Moscone Center, San Francisco , USA
• [Presentation] Treatment of Muscular Dystrophy
  • Author(s): Takeda S
  • Organizer: The 11th Annual Meeting of the Asian and Oceanian Myology Center
  • Place of Presentation: Shirankaikan, Kyoto, Japan
• [Presentation] Gene Therapy for Neuromuscular Disorders
  • Author(s): Takeda S
  • Organizer: Japan Society of Gene Therapy 2012 18th Annual Meeting
  • Place of Presentation: Hotel Kumamoto Terrsa, Kumamoto, Japan