Analysis of splicing variant of p53-inducible phosphatase PPM1D on carcinogenesis

2010 Fiscal Year Final Research Report

• Project/Area Number: 20770095
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Functional biochemistry
• Research Institution: Hokkaido University
• Principal Investigator: CHUMAN Yoshiro Hokkaido University, 大学院・理学研究院, 助教 (40372263)
• Project Period (FY): 2008 – 2010
• Keywords: ホスファターゼ / スプライシング

Research Abstract

PPM1D is a PPM1 type protein phosphatase and is induced in response to DNA damage. In our current study, we identify and characterize an alternative splicing variant (denoted PPM1D430) of human PPM1D at both the mRNA and protein level. PPM1D430 comprises the common 420 residues of the known PPM1D protein (PPM1D605) and contains a stretch of PPM1D430-specific 10 amino acids. Semi-quantitative RT-PCR analysis revealed that PPM1D430 mRNA is also induced in response to the genotoxic stress in a p53-dependent manner. In vitro phosphatase analysis and PPM1D430-specific RNA interference analysis further indicated that PPM1D430 can dephosphorylate Ser15 of human p53 both in vitro and in vivo. On the other hand, expression profiling of this gene by RT-PCR analysis of a human tissue cDNA panel revealed that PPM1D430 is expressed exclusively in testes and in leukocytes whereas PPM1D605 is ubiquitous. In addition, PPM1D430 shows a different subcellular localization pattern and protein stability when compared with PPM1D605 under some conditions. Our current findings thus suggest that PPM1D430 may exert specific functions in immune response and/or spermatogenesis.

Research Products

• [Journal Article] Phosphatase assay for multi-phosphorylated substrates using phosphatase specific-motif antibody (2011)
  • Author(s): Chuman, Y., Iizuka, K., Honda, T., Onoue, H., Shimohigashi, Y., Sakaguchi, K.
  • Journal Title: J.Biochem. in press
  • Peer Reviewed
• [Journal Article] sFRP-1 binds via its netrin-related motif to the N-module of thrombospondin-1 and blocks thrombospondin-1 stimulation of MDA-MB-231 breast carcinoma cell adhesion and migration. (2011)
  • Author(s): Martin-Manso, G., Calzada, M.J., Chuman, Y., Sipes, J.M., Xavier, C.P., Wolf, V., Kuznetsova, S.A., Rubin, J.S., Roberts, D.D.
  • Journal Title: Arch.Biochem.Biophys. 509(2) Pages: 147-156
  • Peer Reviewed
• [Journal Article] Probing phenylalanine environments in oligomeric structures with pentafluorophenylalanine and cyclohexylalanine. (2011)
  • Author(s): Nomura, T., Kamada, R., Ito, I., Sakamoto, K., Chuman, Y., Ishimori, K., Shimohigashi, Y., Sakaguchi, K.
  • Journal Title: Biopolymers 95(6) Pages: 410-419
  • Peer Reviewed
• [Journal Article] Enhancement of transcriptional activity of mutant p53 tumor suppressor protein through stabilization of tetramer formation by calix[6]arene derivatives (2010)
  • Author(s): Kamada, R., Yoshino, W., Nomura, T., Chuman, Y., Imagawa, T., Suzuki, T., Sakaguchi, K.
  • Journal Title: Bioorg.Med.Chem Lett. 20(15) Pages: 4412-4415
  • Peer Reviewed
• [Journal Article] Drastic Effects on Fibril Formation of Amyloid-beta Peptides by the Addition of Amino Acid Residue Units to the Termini (2010)
  • Author(s): Asanomi, Y., Kobayashi, Y, Sakai, H., Masuda, T., Chen, X., Chuman, Y., Uosaki, K., Sakaguchi, K.
  • Journal Title: Protein Pept.Lett. 17(4) Pages: 458-463
  • Peer Reviewed
• [Journal Article] Fluoroalkene Modification of Mercaptoacetamide-based Histone Deacetylase Inhibitors (2010)
  • Author(s): Osada, S., Sano, S., Ueyama, M., Chuman, Y., Kodama, H., Sakaguchi, K.
  • Journal Title: Bioorg.Med.Chem. 18(2) Pages: 605-611
  • Peer Reviewed
• [Journal Article] Characterization of Two Novel and Potent Inhibitors Specific for p53-Inducible Protein Phosphatase PPM1D. (2010)
  • Author(s): Yagi, H., Chuman, Y., Sakaguchi, K.
  • Journal Title: Peptide Science 2009 Pages: 455-456
  • Peer Reviewed
• [Journal Article] Biomineralization Activity of Multivalent minTBP-1 Peptide Mediated by p53 Tetramerization Domain. (2010)
  • Author(s): Sakaguchi, T., Kamada, R., Nomura, T., Chuman, Y., Imagawa, T., Sakaguchi, K.
  • Journal Title: Peptide Science 2009 Pages: 411-412
  • Peer Reviewed
• [Journal Article] Development of Probed-SCAP Directed at Thiol Groups. (2010)
  • Author(s): Watanabe, K., Sakai, H., Chen, X., Chuman, Y, Sakaguchi, K.
  • Journal Title: Peptide Science 2009 Pages: 407-408
  • Peer Reviewed
• [Journal Article] Inhibition of p53 by Introducing Tetramerization Domain Peptide Fused with Polyvalent Cationic Sequence into Cells. (2010)
  • Author(s): Kamada, R., Oshima, S., Chuman, Y., Imagawa, T., Sakaguchi, K.
  • Journal Title: Peptide Science 2009 Pages: 319-320
  • Peer Reviewed
• [Journal Article] PPM1D430, a Novel Alternative Splicing Variant of the Human PPM1D, can Dephosphorylate p53 and Exhibits Specific Tissue Expression (2009)
  • Author(s): Chuman, Y., Kurihashi, W., Mizukami, Y., Nashimoto, T., Yagi, H., Sakaguchi, K.
  • Journal Title: J.Biochem. 145(1) Pages: 1-12
  • Peer Reviewed
• [Journal Article] Effects of tumor-associated mutations in the p53 tetramerization domain on oligomerization state and transcriptional activity. (2009)
  • Author(s): Kamada, R., Terai, T., Nomura, T., Chuman, Y, Imagawa, T., Sakaguchi, K.
  • Journal Title: Adv.Exp.Med.Biol. 611 Pages: 567-568
  • Peer Reviewed
• [Journal Article] Oxidation of methionine residue at hydrophobic core destabilizes p53 tetrameric structure (2009)
  • Author(s): Nomura, T., Kamada, R., Ito, I., Chuman, Y, Shimohigashi, Y, Sakaguchi, K.
  • Journal Title: Biopolymers 91(1) Pages: 78-84
  • Peer Reviewed
• [Journal Article] PPM1D430, a Novel Alternative Splicing Variant of the Human PPM1D. (2009)
  • Author(s): Chuman, Y., Kurihashi, W., Mizukami, Y., Nashimoto, T., Yagi, H., Sakaguchi, K.
  • Journal Title: Peptide Science 2008 Pages: 531-532
  • Peer Reviewed
• [Journal Article] Thermal Stability of p53 Tetramerization Domain Pe4ptides Derived from Various Species. (2009)
  • Author(s): Nomura, T., Kamada, R., Chuman, Y., Sakaguchi, K.
  • Journal Title: Peptide Science 2008 Pages: 35-36
  • Peer Reviewed
• [Journal Article] Identification of Novel Chemical Inhibitors for p53-Inducible Protein Phosphatase PPM1D. (2009)
  • Author(s): Yagi, H., Chuman, Y., Yoshimura, F., Tanino, K., Sakaguchi, K.
  • Journal Title: Peptide Science 2008 Pages: 385-386
  • Peer Reviewed
• [Journal Article] Extremely Long Nanofibrils of Functionalized Structure-Controlled-Amyloid Peptides Derived from Transthyretin. (2009)
  • Author(s): Sakai, H., Asanomi, Y., Kobayashi, Y., Chen, X., Chuman, Y., Masuda, T., Uosaki, K., Sakaguchi, K.
  • Journal Title: Peptide Science 2008 Pages: 465-466
  • Peer Reviewed
• [Journal Article] Characterization of the Active Site and a Unique Uncompetitive Inhibitor of the PPM1-type Protein Phosphatase PPM1D (2008)
  • Author(s): Chuman, Y., Yagi, H., Fukuda, T., Nomura, T., Matsukizono, M., Shimohigashi, Y., Sakaguchi K.
  • Journal Title: Protein Pept.Lett. 15 Pages: 938-948
  • Peer Reviewed
• [Journal Article] Differential Recognition of Phosphorylated Transactivation Domains of p53 by Different p300 Domains (2008)
  • Author(s): Polley, S., Guha, S., Roy, N.S., Kar, S., Sakaguchi, K., Chuman, Y., Swaminathan, V., Roy, S.
  • Journal Title: J.Mol.Biol. 376 Pages: 8-12
  • Peer Reviewed
• [Journal Article] Correlation of Oligomeric Structure and Transcriptional Activity of p53 Mutants. (2008)
  • Author(s): Kamada, R, Terai, T., Nomura, T., Chuman, Y, Imagawa, T., Sakaguchi, K.
  • Journal Title: Peptide Science 2007 Pages: 69-70
  • Peer Reviewed
• [Journal Article] Screening of Peptides to Stabilize the p53 Tetramer Formation from Phage Displayed Library. (2008)
  • Author(s): Nomura, T., Kamada, R., Chuman, Y., Sakaguchi, K.
  • Journal Title: Peptide Science 2007 Pages: 315-316
  • Peer Reviewed
• [Journal Article] Correlation between Expression Level in E. coli and Stability of Oligomeric Structure of the p53 Tetramerization Domain. (2008)
  • Author(s): Miyazaki, H., Kamada, R., Nomura, T., Chuman, Y., Imagawa, T., Sakaguchi, K.
  • Journal Title: Peptide Science 2007 Pages: 317-318
  • Peer Reviewed
• [Journal Article] Structural Photomodulation of Peptides with Azobenzoate Derivative in Peptide Backbone. (2008)
  • Author(s): Kamada, R., Fukuda, K., Miyazaki, H., Nomura, T., Chuman, Y., Imagawa, T., Tanino, K., Sakaguchi, K.
  • Journal Title: Peptide Science 2007 Pages: 447-448
  • Peer Reviewed
• [Presentation] Novel chemical inhibitors specific for p53-inducible protein phosphatase PPM1D (2011)
  • Author(s): Yagi Y.
  • Organizer: Anaheim Convention Center, Anaheim, USA
  • Place of Presentation: Anaheim Convention Center, Anaheim, USA
  • Year and Date: 2011-03-29
• [Presentation] Suppression of Breast Cancer Cells Proliferation by Novel Inhibitor for p53-Inducible Protein Phosphatase PPM1D (2011)
  • Author(s): Chuman Y.
  • Organizer: 1st International Symposium on Carcinogenic Spiral & 9th International Conference on Protein Phosphatase
  • Place of Presentation: Tetsumon Memorial Hall, The University of Tokyo
  • Year and Date: 2011-02-01
• [Presentation] Novel small molecule inhibitors specific for p53-inducible protein phosphatase PPM1D (2011)
  • Author(s): Yagi H.
  • Organizer: 1st International Symposium on Carcinogenic Spiral & 9th International Conference on Protein Phosphatase
  • Place of Presentation: Tetsumon Memorial Hall, The University of Tokyo
  • Year and Date: 2011-02-01
• [Presentation] Increase in number of nucleoli by protein phosphatase PPM1D overexpression (2011)
  • Author(s): Kozakai Y.
  • Organizer: 1st International Symposium on Carcinogenic Spiral & 9th International Conference on Protein Phosphatase
  • Place of Presentation: Tetsumon Memorial Hall, The University of Tokyo
  • Year and Date: 2011-02-01
• [Presentation] p53誘導性ホスファターゼPPMID阻害による乳がん細胞増殖抑制作用 (2010)
  • Author(s): 中馬吉郎
  • Organizer: BMB2010第83回日本生化学会、第33回日本分子生物学会合同大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-10
• [Presentation] p53誘導性ホスファターゼPPMID特異的阻害剤SPI-001の相互作用 (2010)
  • Author(s): 八木寛陽
  • Organizer: BMB2010第83回日本生化学会、第33回日本分子生物学会合同大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-10
• [Presentation] PPMID430, a Nove1 Alternative Splicing Variant of the Human PPM1D, can Dephosphorylate p53 and Exhibits Specific Tissue Expression (2010)
  • Author(s): 中馬吉郎
  • Organizer: BMB2010第83回日本生化学会第33回日本分子生物学会合同大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-09
• [Presentation] Dephosphorylation of Nucleolar Protein Nucleophosmin by Protein Phosphatase PPM1D (2010)
  • Author(s): Kozakai Y.
  • Organizer: 5th International Peptide Symposium in conjunction with 47th Japanese Peptide Symposium
  • Place of Presentation: Kyoto International Conference Center
  • Year and Date: 2010-12-04
• [Presentation] Effect of Protein Phosphatase PPM1D on Nucleolus Formation (2010)
  • Author(s): Kozakai Y.
  • Organizer: MPSA 2010
  • Place of Presentation: Norrlands nation at Uppsala University in Uppsala, Sweden
  • Year and Date: 2010-08-25
• [Presentation] p53誘導性ホスファターゼ特異的阻害剤の開発と細胞増殖抑制能 (2010)
  • Author(s): 中馬吉郎
  • Organizer: 日本化学会北海道支部2010年夏季研究発表会
  • Place of Presentation: 函館高専
  • Year and Date: 2010-07-24
• [Presentation] p53誘導性ホスファターゼのスプライシングバリアントの同定と阻害剤開発 (2010)
  • Author(s): 中馬吉郎
  • Organizer: 第47回日本生化学会北海道支部例会(招待講演)
  • Place of Presentation: 北海道大学医学部第3講堂
  • Year and Date: 2010-07-23
• [Presentation] p53誘導性ホスファターゼPPMID阻害剤における特定に配向する疎水性部位 (2010)
  • Author(s): 八木寛陽
  • Organizer: 第47回日本生化学会北海道支部例会
  • Place of Presentation: 北海道大学医学部第3講堂
  • Year and Date: 2010-07-23
• [Presentation] PPMIDホスファターゼ過剰発現が核小体形成に及ぼす影響 (2010)
  • Author(s): 小境夕紀
  • Organizer: 第47回日本生化学会北海道支部例会
  • Place of Presentation: 北海道大学医学部第3講堂
  • Year and Date: 2010-07-23
• [Presentation] 脱リン酸化酵素PPMID阻害剤SPI-001の阻害様式と核小体形成に対する効果 (2010)
  • Author(s): 小境夕紀
  • Organizer: 日本化学会第91春季年会(2011)
  • Place of Presentation: 神奈川大学横浜キャンパス
  • Year and Date: 2010-03-29
• [Presentation] p53誘導性ホスファターゼPPMIDの新規阻害剤 (2010)
  • Author(s): 梨本健紘
  • Organizer: 日本薬学会第130年会
  • Place of Presentation: 岡山桃太郎アリーナ(岡山)
  • Year and Date: 2010-03-29
• [Presentation] p53誘導性ホスファターゼPPMIDに対する新規阻害剤の開発 (2009)
  • Author(s): 中馬吉郎
  • Organizer: 第4回日本プロテインホスファターゼ研究会学術集会
  • Place of Presentation: 熊本大学(熊本)
  • Year and Date: 2009-11-13
• [Presentation] Characterization of Two Novel and Potent Inhibitors Specific for p53-Inducible Protein Phosphatase PPM1D (2009)
  • Author(s): Yagi H.
  • Organizer: 第46回ペプチド討論会
  • Place of Presentation: 北九州国際会議場(小倉・北九州)
  • Year and Date: 2009-11-04
• [Presentation] p53誘導性ホスファターゼPPMIDの基質認識と阻害剤開発 (2009)
  • Author(s): 八木寛陽
  • Organizer: 第42回若手ペプチド夏の勉強会
  • Place of Presentation: 八王子セミナーハウス(東京・八王子)
  • Year and Date: 2009-08-02
• [Presentation] p53誘導性ホスファターゼPPMIDに対するシリル基を持つ低分子阻害剤 (2009)
  • Author(s): 八木寛陽
  • Organizer: 日本生化学会北海道支部第46回例会
  • Place of Presentation: 旭川医科大学(旭川)
  • Year and Date: 2009-07-17
• [Presentation] Identification and characterization of novel alternative splicing variant of p53-inducible protein phosphatase PPMID (2008)
  • Author(s): Kurihashi W.
  • Organizer: 第8回ホスファターゼカンファレンス
  • Place of Presentation: 前橋テルサ(群馬)
  • Year and Date: 2008-11-14
• [Presentation] ヒトPPMIDの選択的スプライシングバリアントPPMID430 (2008)
  • Author(s): 中馬吉郎
  • Organizer: 第45回ぺプチド討論会
  • Place of Presentation: タワーホール船堀(東京都江戸川区)
  • Year and Date: 2008-10-31
• [Presentation] Identification of novel chemical inhibitors for p53-inducible protein phosphatase PPMID (2008)
  • Author(s): 八木寛陽
  • Organizer: 第45回ペプチド討論会
  • Place of Presentation: タワーホール船堀(東京都江戸川区)
  • Year and Date: 2008-10-31
• [Presentation] An Alternative Splicing Variant of the PPM1 Protein Phosphatase PPMID (2008)
  • Author(s): Chuman Y.
  • Organizer: MPSA2008
  • Place of Presentation: 北海道大学(札幌市)
  • Year and Date: 2008-08-26
• [Presentation] Novel chemical inhibitors for p53-inducible protein phosphatase PPM1D (2008)
  • Author(s): Yagi H.
  • Organizer: MPSA2008
  • Place of Presentation: 北海道大学(札幌市)
  • Year and Date: 2008-08-26
• [Presentation] リン酸化と脱リン酸化によるタンパク質制御機構～p53誘導性ホスファターゼPPMIDを中心にして～ (2008)
  • Author(s): 中馬吉郎
  • Organizer: 第41回若手ペプチド夏の勉強会
  • Place of Presentation: プライムイン城陽(京都・城陽市)
  • Year and Date: 2008-08-05
• [Presentation] 癌抑制タンパク質p53誘導性ホスファターゼのAlternativesplicing variant (2008)
  • Author(s): 中馬吉郎
  • Organizer: 日本分子生物学会第8回春季シンポジゥム
  • Place of Presentation: 京王プラザホテル(札幌)
  • Year and Date: 2008-05-27
• [Patent(Industrial Property Rights)] プロテインホスファターゼ阻害剤 (2008)
  • Inventor(s): 坂口和靖、谷野圭持、中馬吉郎、吉村文彦、八木寛陽
  • Industrial Property Rights Holder: LCIP
  • Industrial Property Number: 特許,特願2008-262646,PCT出願PCT/JP2009/005128,国際公開番号:WO2010041401A1
  • Filing Date: 2008-10-09