• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Direct Observation of the Regulation of Microtubule Dynamics by MAP2 Phosporylation

Research Project

Project/Area Number 07680715
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionNagoya University

Principal Investigator

ITOH Tomohiko  Nagoya University Graduate School of Science Associate Professor, 大学院・理学研究科, 講師 (30183742)

Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1996: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1995: ¥1,500,000 (Direct Cost: ¥1,500,000)
Keywordsmicrotubule / dynamic instability / MAP2 / phosphorylation / dark-field microscopy / クラスター形成 / 蛍光標識 / 光学顕微鏡観察 / 微小管の動態 / cdc2 Kinase
Research Abstract

Phosphorylation-dependent regulation of microtubule-stabilizing activities of microtubule-associated protein 2 (MAP2) was examined using optical microscopy. MAP2, purified from mammalian brain, was phosphorylated by either cAMP-dependent protein kinase (PKA) or cyclin B-dependent cdc2 kinase. By PKA,15 mloles of phoshoryl groups was incorporated into one mole of MAP2, but about 70% was distributed to the projection region. By cdc2 kinase, 8-10 moles of phosphoryl groups was incorporated into one mole of MAP2 and more than 60% of the phosphates was distributed to the microtubule-binding region. Both phosphorylations similarly reduced binding activity of MAP2 onto microtubules. Direct observation of individual microtubules under a dark-field microscope showed that interconversion between polymerization phase and depolymerization phase was repeated in both unphosphorylated and PKA-phosphorylated MAP2. In cdc2 kinase-phosphorylated MAP2, however, phase transition from depolymerization to polymerization was difficult to take place, with the result that life-time of individual microtubules was as short as in the absence of MAP2. Examination of spontaneous nucleation of microtubules under a dark-field microscope showed that PKA-phosphorylated MAP2 redued nucleating-activity, while cdc2 kinase-phosphorylated MAP2 completely abolished it. These observations show that cdc2 kinase-dependent phosphorylation inhibited both microtubule-stabilizing activity and microtubule-nucleating activity of MAP2, while PKA-dependent phosphorylation affected only microtubule-nucleating activity of MAP2.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] K. Ookata: "Cyclin B interaction with microtubule-associated protein 4 (MAP4) targets p34^<COC2> kinase to microtubules and is potential regulator of M-phase microtubute dynamics" Journal of Cell Biology. 128 (5). 849-862 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T. J. ltoh: "Phosphorylation States of Microtubule-Associated Protein 2 (MAP2) Determine the Regulatory Role of MAP2 in Microtubule Dynamics" Biochemistry. 33(発表予定). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Ookata, K.: "Cyclin B Interaction with Microtubule-Associated Protein 4 (MAP4) Targets P34^<CDC2> Kinase to Microtubules and Is a Potential Regulator of M-Phase Microtubule Dynamics." J.Cell Biol.128(5). 849-862 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Itoh, T.J.: "Phosphorylation States of Microtubule-Associated Protein 2 (MAP2) Determine the Regulatory Role of MAP2 in Microtubule Dynamics." Biochemistry. (in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.J.Itoh: "Phosphorylation States of Microtubule-Associated Protein 2 (MAP2) Determine the Regulatory Role of MAP2 in Microtubule Dynamics" Biochemistry. 33(発表予定). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Ookata: "Cyclin B interaction with microtubule-associated protein 4(MAP4)targets p34^<cdc2> Kinase to microtubules and is potential regulator of M-phase microtubule dynamics" Journal of Cell Biology. 128. 849-862 (1995)

    • Related Report
      1995 Annual Research Report

URL: 

Published: 1995-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi