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Analysis of cross-bridge structures between the nuclear membrane and cytoplasmic microtubules.

Research Project

Project/Area Number 05804051
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 生物形態・構造
Research InstitutionThe Tokyo Metropolitan Institute of Medical Science

Principal Investigator

SAMESHIMA Masazumi  The Tokyo Metropolitan Institute of Medical Science, Department of U1trastructural Research, Research Scientist., 超微形態研究部門, 研究員 (90142653)

Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1994: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1993: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsDictyostelium discoideum / Microtubules / Nucleus / 微小菅
Research Abstract

At the aggregation stage of Dictyostelium discoideum, the signal transduction system depending on cAMP becomes active. We recently have found that cross-bride structures are formed between the outer nuclear membrane and cytoplasmic microtubules (MTs) at this stage. To search molecules composing these structures, microtubule-associated proteins (MAPs) of D.discoideum were examined.Although there were no bands specific to aggregating cells, several common components, 42,52,55,59,72,75,95,220 and 280 kDa bands, were detected on SDS-PAGE.Each component was analyzed with western blotting and their partial amino acid sequences also were examined.
(1) The 55 kDa band is originated from depolymerized porcine tubulin.
(2) The 42 and 95 kDa are actin and an actin-cross linking protein, alpha actinin, respectively. Partial amino acid sequence of the 75 kDa component is similar to HSC70, actin-capping protein.
(3) MTs fraction incubated with Dictyostelium cell sup has been examined by immuno-negative staining. Signals for anti-actin antibody are found on structures attached to the terminal end of MTs. If this is the+end, this type of binding of actin to MTs is interested from a point of view of the relation of MTs and actin systems under the cytoplasmic membranes.
(4) Partial amino acid sequence of the 280 kDa band coincides with human cytokeratin 9. Since this band reproducibly is found and molecular weight of the keratin is 64 kDa, this band seems not to be originated simply from contaminations of human debris during experiments.
(5) Little is known about other bands. It will be possible to determine molecules composing cross-bridge structures through the examination of intracellular distribution using antibody against each band separated on the gels

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (11 results)

All Other

All Publications (11 results)

  • [Publications] M.Sameshima: "Disappearance of the surface sheath from Dictyostelium discoideum sorus." Cytologia. 58. 361-366 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Sameshima: "Novel actin rods appeared in spores of Dictyostelium discoideum." Cell Struct.Funct.19. 189-194 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kishi: "Differential interference microscopy of intranuclear actinrods in Dictyostelium discoideum spores." Cytologia. 59. 453-460 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Sudha: "Abnormal integrity of nucleolus associated with cell cycle arrest due to the temperature-sensitive ubiquitin-activatingenzyme E1." Chromosome Research. 3. 115-123 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Sameshita, M: "Disappearance of the surface sheath from Dictyostelim discoideumsorus." Cytologia.58. 361-366 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Sameshita, M., et al.: "Novel actin rods appeared in spores of Dictyostelium discoideum." Cell Struct.Funct.19. 189-194 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kishi, Y., et al.: "Differential interference microscopy of intranuclear actin rods in Dictyostelium discoideum." Cytologia.59. 453-460 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Sudha, T., et al.: "Abnormal integrity of nucleolus associated with cell cycle arrest due to the temperarure-sensitive ubiquitin-activating enzyme E1." Chromosome Res.3. 115-123 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kishi: "Differential interference microscopy of intranuclear actin rods in Dictyostelium discoideum spores." Cytologia. 59. 453-460 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] T.Sudha: "Abnormal integrity of nucleolus associated with cell cycle arrest due to the temperature-sensitive ubiquitin-activating enzyme El." Chromosome Research. in press.

    • Related Report
      1994 Annual Research Report
  • [Publications] M.Sameshima: "Disappearance of the surface sheath from Dictyostelium discoideum sorus during late culmination." Cytologia. 58. 361-366 (1993)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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