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Global identifications and analyses of the proteins that interact with chitin synthases of filamentous fungi.

Research Project

Project/Area Number 15380057
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied microbiology
Research InstitutionThe University of Tokyo

Principal Investigator

HORIUCHI Hiroyuki  The University of Tokyo, Graduate School of Agricultural and Life Sciences, Associate Professor, 大学院・農学生命科学研究科, 助教授 (00209280)

Project Period (FY) 2003 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥15,600,000 (Direct Cost: ¥15,600,000)
Fiscal Year 2004: ¥7,500,000 (Direct Cost: ¥7,500,000)
Fiscal Year 2003: ¥8,100,000 (Direct Cost: ¥8,100,000)
Keywordschitin synthase / Aspergillus / myosin / septum / actin
Research Abstract

We have cloned five chitin synthase genes from filamentous fungus Aspergillus nidulans and investigated their functions. In this project, we constructed the strains that produced functional epitope-tagged chitin synthases to clarify the localization of their gene products. HA epitope-tagged ChsA (HA-ChsA) mainly localized to forming septa, while GFP or FLAG epitope-tagged ChsB (GFP-ChsB or FLAG-ChsB) and FLAG epitope-tagged ChsC (FLAG-ChsC) mainly localized to forming septa and tips of hyphae. We next constructed the strain that produced both HA-ChsA and FLAG-ChsC and investigated their co-localization. The localizations of HA-ChsA and FLAG-ChsC were partly overlapped. Western blot analyses against the cell lysates of the strains expressing HA-ChsA, FLAG-ChsC, and FLAG-ChsB showed that each of them was present as several bands in the cell, suggesting that they underwent post-transcriptional modifications. We showed by phosphatase treatment that a portion of FLAG-ChsB were phosphorylate … More d. We also constructed chsA chsC chsD and chsA chsC csmA triple deletion mutants and indicated that csmA played crucial role(s) for the growth of the chsA chsC double mutant.
Next, we constructed the strains that expressed the CsmA tagged with nine copies of HA eppitope (CsmA-HA) instead of wild-type CsmA. CsmA-HA localized at hyphal tips and forming sepia. We showed that the myosin motor-like domain of CsmA was able to bind to actin in vivo and in vitro and that this binding is indispensable for the correct localization and function of CsmA. We isolated a paralog of csmA, csmB, from A. nidulans and characterized its function. Growth and morphological defects of csmB deletion mutants were partly similar to those of the csmA deletion mutant. We showed that deletions of csmA and csmB were synthetically lethal. We next constructed the strains that produced both CsmA-HA and CsmB tagged with three copies of FLAG epitope at its C-terminus and investigated their co-localization. Their localizations were partly overlapped. Less

Report

(3 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • Research Products

    (7 results)

All 2005

All Journal Article (7 results)

  • [Journal Article] The class V chitin synthase gene csmA is crucial for the growth of the chsA chsC double mutant in Aspergillus nidulans.2005

    • Author(s)
      山田絵美, 一宮維幸, 太田明徳, 堀内裕之
    • Journal Title

      Biosci.Biotechnol.Biochem. 69

      Pages: 87-97

    • NAID

      130000030558

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] CsmA, a class V chitin synthase with a myosin motor-like domain, is localized through direct interaction with the actin cytoskeleton in Aspergillus nidulans.2005

    • Author(s)
      竹下典男, 太田明徳, 堀内裕之
    • Journal Title

      Mol.Biol.Cell 16

      Pages: 1961-1970

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Class I and class II chitin synthases are involved in septum formation in the filamentous fungus Aspergillus nidulans.2005

    • Author(s)
      一宮維幸, 山田絵美, 山下修一, 太田明徳, 堀内裕之
    • Journal Title

      Eukaryotic Cell 4(印刷中)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The class V chitin synthase gene csmA is crucial for the growth of the chsA chsC double mutant in Aspergillus nidulans.2005

    • Author(s)
      Emi Yamada, Masayuki Ichinomiya, Akinori Ohta, Hiroyuki Horiuchi
    • Journal Title

      Biosci.Biotechnol.Biochem. 69

      Pages: 87-97

    • NAID

      130000030558

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] CsmA, a class V chitin synthase with a myosin motor-like domain, is localized through direct interaction with the actin cytoskeleton in Aspergillus nidulans.2005

    • Author(s)
      Norio Takeshita, Akinori Ohta, Hiroyuki Horiuchi
    • Journal Title

      Mol.Biol.Cell 16

      Pages: 1961-1970

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Class I and class II chitin synthases are involved in septum formation in the filamentous fungus Aspergillus nidulans.2005

    • Author(s)
      Masayuki Ichinomiya, Emi Yamada, Shuichi Yamashita, Akinori Ohta, Hiroyuki Horiuchi
    • Journal Title

      Eukaryotic Cell 4 (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] CsmA, a class V chitin synthase with a myosin motor-like domain, is localized through direct interaction with the actin cytoskeleton in Aspergillus nidulans.2005

    • Author(s)
      竹下典男, 太田明徳, 堀内裕之
    • Journal Title

      Mol.Biol.Cell 16(印刷中)

    • Related Report
      2004 Annual Research Report

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

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