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Structural and functional analysis on protein complex of ubiquitin-proteasome system

Research Project

Project/Area Number 15580308
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied molecular and cellular biology
Research InstitutionYokohama City University

Principal Investigator

KAWASAKI Hiroshi  Yokohama City University, International Graduate School of Arts and Sciences, Associate Professor, 国際総合科学研究科, 准教授 (70169704)

Co-Investigator(Kenkyū-buntansha) SASSA Hidenori  Chiba university, Fucuity of Horticulture, Associate Professor, 園芸学部, 准教授 (50295507)
Project Period (FY) 2003 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2006: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2005: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2004: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2003: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsProtein / Proteomics / Biotechnology / Proteasome / Ubiquitin / Protein complex / Posttranslational modification / 蛋白質 / プロテオーム / 生態機能利用 / TAP法
Research Abstract

Protein degradation by ubiquitin-proteasome system is a major path of protein catabolism in cells. The ubiquitin-proteasome system regulates cell cycle, antigen presentation and plant self compatibility.
We have isolated proteasome and protein complex interacting proteasome subunits, and analyzed structure and functions. Four rice ATPase subunits of the proteasome were encoded by duplicated genes. We purified rice 19S complex of the proteasome and analyzed the expression of each gene products in various rice tissues. Tissue specific expression of each product suggests the preesence of multiform 19S complex engaged in the tissue-specific protein metabolism. The posttranslational modification of yeast 20S proteasome was analyzed. The phosphorylation sites of alpha7 subunit was determined. RPN4, originally identified as a subunit of 26S proteasome, is a transcriptional regulator for several subunits of proteasome. We isolated RPN4 complex from yeast cells by TAP method. Bud32 is reported to interact many subunits of proteasome. We isolated Bud32 complex from yeast haploid cells. The complex consisted of four proteins, Kael, Cgi121, Bud32 and Gon7. This complex was reported recently as KEOPS (for kinase, putative endopeptidase and other proteins of small size)/EKC (Endopeptidase-like Kinase Chromatin-associated), which regulates telomere maintenance or transcription of several genes. These proteins observed in nucleus and also in cytosol. The duploid deletion mutant of Bud32 shows random budding instead of bipolar budding pattern observed in normal cells. Budding pattern of deletion mutants on other members of the complex was not random. Bud32 complex isolated from duploid cells contained similar proteins. Some additional minor components were observed. These results suggest that Bud32 itself regulates budding pattern of duploid yeast cells.

Report

(5 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • Research Products

    (33 results)

All 2006 2005 2004 2003 Other

All Journal Article (26 results) Book (4 results) Publications (3 results)

  • [Journal Article] Identification of yeast aspartylaminopeptidase gene by purifying and characterizing its product from yeast cells.2006

    • Author(s)
      Yokoyama, R.
    • Journal Title

      FEBS J. 273

      Pages: 192-198

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary 2005 Annual Research Report
  • [Journal Article] A novel approach and protocol for discovering extremely low-abundance proteins in serum2006

    • Author(s)
      Tanaka, Y.
    • Journal Title

      Proteomics 6

      Pages: 4845-4888

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Proteomic search for potential diagnostic markers and therapeutic targets for ovarian clear cell adenocarcinoma2006

    • Author(s)
      Morita, A.
    • Journal Title

      Proteomics 6

      Pages: 5880-5890

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Efficient electroblotting of gel-resolved proteins onto diamond-like carbon-coated plate for protein-chip2006

    • Author(s)
      Ino, Y.
    • Journal Title

      J. Electrophoresis 50

      Pages: 33-37

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Identification of yeast aspartylaminopeptidase gene by purifying and characterizing its product from yeast cells2006

    • Author(s)
      Yokoyama, R.
    • Journal Title

      FEBS J. 273

      Pages: 192-198

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Proteomic search for potential diagnostic markers and therapeutic targets for ovarian clear dell adenocarcinoma2006

    • Author(s)
      Morita, A.
    • Journal Title

      Proteomics 6

      Pages: 5880-5890

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Efficient electroblotting og gel-resolved proteins onto daimond-like carbon-coated plate for protein-chip2006

    • Author(s)
      Ino, Y.
    • Journal Title

      J.Electrophoresis 50

      Pages: 33-37

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] A novel approach and protocol for discovering extremely low-abundance proteins in serum2006

    • Author(s)
      Y.Tanaka
    • Journal Title

      Proteomics 6

      Pages: 4845-4855

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Proteomic search for potential diagnostic markers and therapeutic targets for ovarian clear cell adenocarcinoma2006

    • Author(s)
      A.Morita
    • Journal Title

      Proteomics 6

      Pages: 5880-5890

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Efficient electroblotting of gel-resolved proteins onto diamond-like carbon-coated plate for protein-chip2006

    • Author(s)
      Y.Ino
    • Journal Title

      J. Electrophoresis 50

      Pages: 33-37

    • Related Report
      2006 Annual Research Report
  • [Journal Article] プロテオミクスで見えてくる翻訳後修飾の機能2005

    • Author(s)
      川崎博史
    • Journal Title

      実験医学 23(7)

      Pages: 1043-1049

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary 2004 Annual Research Report
  • [Journal Article] Assessment of filter plates for multi-well in-gel digestion of proteins separated by polyacrylamide gel electrophoresis to identify them with LC-ESI/MSMS.2005

    • Author(s)
      Kameyama, K.
    • Journal Title

      J Electrophoresis 49

      Pages: 71-75

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary 2005 Annual Research Report
  • [Journal Article] Functional analysis of posttranslational modification by proteomics2005

    • Author(s)
      Kawasaki, H.
    • Journal Title

      Jikkenigaku 23

      Pages: 1043-1049

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Assesment of filter plates for multi-well in-gel digestion of proteins of proteins separated by polyacrylamide gel electrophoresis to identify them with LC-ESI/MSMS2005

    • Author(s)
      Kameyama, K.
    • Journal Title

      J.Electrophoresis 49

      Pages: 71-75

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Identification of three phosphorylation sites in the α 7 subunit of the yeat 20S proteasome in vivo using mass spectrometry2004

    • Author(s)
      Iwafune, Y.
    • Journal Title

      Arch Biochem Biophys. 431

      Pages: 9-15

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Identification of three phosphorylation sites in the alpha7 subunit of the yeast 20S proteasome in vivo using mass spectrometry2004

    • Author(s)
      Iwafune, Y.
    • Journal Title

      Arch.Biochem.Biophys. 431

      Pages: 9-15

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Parvalbumin2004

    • Author(s)
      Nakayama, S.
    • Journal Title

      Handbook of Metalloproteins Vol. 3, JohnWiley & Sons

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Sample preparation2004

    • Author(s)
      Kawasaki, H.
    • Journal Title

      Genomics, proteomics no sintennkai(NTS)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Sequnce determination2004

    • Author(s)
      Hirano, H.
    • Journal Title

      Genomics, proteomics no sintennkai(NTS)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Identification of three phosphorylation sites in the α7 subunit of the yeat 20S proteasome in vivo using mass spectrometry2004

    • Author(s)
      Iwafune, Y.
    • Journal Title

      Arch Biochem Biophys 431

      Pages: 9-15

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Mass spectrometric analysis of expression of ATPase subunits encoded by duplicated genes in the 19S regulatory particle of rice 20S proteasome2003

    • Author(s)
      Shibahara, T.
    • Journal Title

      Arch. Biochem. Biophys. 421

      Pages: 34-41

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] 翻訳後修飾の解析2003

    • Author(s)
      川崎博史
    • Journal Title

      臨床検査 47(11)

      Pages: 1233-1240

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] 26Sプロテアソームサブユニットの翻訳後修飾2003

    • Author(s)
      岩船裕子
    • Journal Title

      生物物理化学 47(4)

      Pages: 143-149

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Mass spectrometric analysis of expression of ATPase subunits encoded by duplicated genes in the 19S regulatory partilce of rice 20S proteasome2003

    • Author(s)
      Shibahara, T.
    • Journal Title

      Arch.Biochem.Biophys. 421

      Pages: 34-41

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Analysis of posttranslational modifactions2003

    • Author(s)
      Kawasaki, H.
    • Journal Title

      Rinshokensa 47

      Pages: 1233-1240

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Posttranslational modifications of the subunits of 20S proteasome2003

    • Author(s)
      Iwafune, Y.
    • Journal Title

      Seibutubuturikagaku 47

      Pages: 143-149

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Book] Parvalbumin. Handbook of Metalloproteins. Vol. 32004

    • Author(s)
      Nakayama, S.
    • Publisher
      John Wiley & Sons, Ltd
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Book] 試料前処理法,ゲノミクス・プロテオミクスの新展開2004

    • Author(s)
      川崎博史
    • Publisher
      エヌ・ティー・エス
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary 2004 Annual Research Report
  • [Book] 一次構造決定,ゲノミクス・プロテオミクスの新展開2004

    • Author(s)
      平野久
    • Publisher
      エヌ・ティー・エス
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary 2004 Annual Research Report
  • [Book] Parvalbumin., Handbook of Metalloproteins.Vol.32004

    • Author(s)
      Nakayama, S.
    • Total Pages
      8
    • Publisher
      John Wiley & Sons, Ltd
    • Related Report
      2004 Annual Research Report
  • [Publications] Shibahara, T.: "Mass spectrometric analysis of expression of ATPase subunits encoded by duplicated genes in the 19S regulatory particle of rice 26S proteasome"Arch.Biochem.Biophys.. 421. 34-41 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 川崎博史: "翻訳後修飾の解析"臨床検査. 47(11). 1233-1240 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 岩船裕子: "26Sプロテアソームサブユニットの翻訳後修飾"生物物理化学. 47(4). 143-149 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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