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Molecular biological char acterization of regulatory mechanism of gene expression in filamentous fungi

Research Project

Project/Area Number 03453138
Research Category

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

Allocation TypeSingle-year Grants
Research Field 応用微生物学・発酵学
Research InstitutionNAGOYA UNIVERSITY

Principal Investigator

TSUKAGOSHI Norihiro  NAGOYA UNIVERSITY,DEPT.of AGRICULTURE, PROFESSOR, 農学部, 教授 (50115599)

Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1992: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1991: ¥5,000,000 (Direct Cost: ¥5,000,000)
KeywordsAspergillus oryzae / Taka-amylase A gene / Trans-factor / AnCP1 / AnNP1 / CCAAT-box / トランスファクター / CCAAT-box / タカアミラ-ゼA遺伝子
Research Abstract

Taka-amylase A (Taa), one of alpha -amylases,is inducibly synthesized and secreated by Aspergillus oryzae. We investigated the expression regulatory mechanism of the Taa gene from Aspergill us oryzae by using Aspergillus nidulans as an intermediate host. The induction of Taa was shown to be regulated at the transcription level by analyzing the transcripts specific for Taa gene synthesize dinnuclei isolated from starch-and glucose-grown cells, A. nidulans carrying the A.oryzae Taa gene. Taa gene specific transcript was detected only in the nucl ei isolated from cells grown under the inducible conditions, namely starch-grown cells. Potential candidates for sequence elements essential for starch inducibility are present upstream of the transcription start point;a CCAAT box, a GC box, an octermer motif-like sequence. Full inducibility was preserved up to deletion-362,as long as the CCAAT sequence was intact. Further analysis showed that a 55bpDNA fragment containing the CCAAT sequence was long enough to confer starch inducibility on the Taa gene. Nuclear extracts from starch-and glucose-grown cells were assayed for proteins which bind to the promoter region of the Taa gene. A protein designated AnCP 1 was detected in nuclear extracts of both starch-and glucose-grown cells and was found to bind to the CCAAT sequence. Unexpectedly,the other DNA binding protein designated AnNP1 was detected in the nuclear extract of glucose grown cells and was found to bind to the 25bp region just upstream of the AnCP1 binding site. Occupancy of the two binding sites appeared to be mutually exclusive, which is suggestive of a negative regulatory mechanism for Taa gene expression.

Report

(3 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] O.Nagata等: "Aspergillus nidulans naclear proteins bind to a CCAAT element and the adjacent upstream seguence in the promoter region of the starch-inducible Taka-amylase A gene" Molecular and General Genetics. (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] S.Ebisu等: "Production of a jungal protein,Taka-amylase A,by protein-producing Bacillas brevis HPD31" Journal of Industrial Microbiology. (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] O. Nagata et al.: "Aspergillus nidulans nuclear proteins bind to a CCAAT element and the adjacent upstream sequence in the promoter region of the starch-inducible Taka-amylase A gene" Molecular and General Genetics. (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] S. Ebisu et al.: "Production of a fungal protein, Taka-amylase A, by protein-producing Bacillus brevis HPD31" Journal of Industrial Microbiology. (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] O.Nagata 等: "Aspergillus nidulans nuclear proteins bind to a CCAAT element and the adjacent upstream seguence in the promoter region of the stanch-inducible Taka-amylase A gene" Molecular and General Genefics. (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Ebisu 等: "Production of a fungal protein,Taka-amylase A,by protein-producing Bacillus brevis HPD 31" Journal of Industrial Microbiology. (1993)

    • Related Report
      1992 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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