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Isolation and characterization of a dnaK genomic locus in a halotolerant cyanobacterium Aphanothece halophytica

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Abstract

We cloned and characterized a genomic locus encoding a distinct member of the DnaK/Hsp70 family of molecular chaperones, dnaK1, from the halotolerant cyanobacterium Aphanothece halophytica. Co-expression of dnaK1 with a plant plastocyanin precursor in Escherichia coli resulted in a dramatic increase in the solubility of the plant protein. This indicates that A. halophytica dnaK1 encodes a functional protein possessing functions assigned to DnaK/Hsp70 chaperone members. The A. halophytica dnaK1 locus also encompasses grpE and dnaJ homologue genes in the order grpE-dnaK1-dnaJ. The transcript content of dnaK1 increased strongly upon subjecting cyanobacterial cells to heat stress. Northern analyses using specific probes indicated transcript species of 2.8, 2.2, 1.3, and 0.7 kb, which comprised grpE-dnaK1, dnaK1, dnaJ, and grpE, respectively. This indicates the presence of different terminators and/or heat stress promoters in this locus. Both dnaK1 transcript and protein levels increased in cyanobacterial cells transferred to hyperosmotic environments, suggesting a role of DnaK1 in the protection and/or recovery of A. halophytica from this particular stress.

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Authors and Affiliations

  1. BioScience Center, School of Agricultural Science, Nagoya University, Chikusa-ku, Nagoya, Aichi, 464–01, Japan

    Byung Hyun Lee

  2. Department of Chemistry, Faculty of Science & Technology, Meijo University, 1–510 Shiogamaguchi, Tenpaku-ku, Nagoya, Aichi, 468, Japan

    Takashi Hibino & Teruhiro Takabe

  3. Department of Dairy Science, College of Agriculture, Kyungpook National University, Taege, 702–701, Korea

    Jinki Jo

  4. PROMUBIE (CONICET), Departamento de Microbiologia, Facultad de Ciencias Bioquimicas y Farmaceuticas, Universidad Nacional de Rosario, Suipacha 531, 2000, Rosario, Argentina

    Alejandro M. Viale

  5. Research Institute of Meijo University, Tenpaku-ku, Nagoya, Aichi, 468, Japan

    Teruhiro Takabe

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  1. Byung Hyun Lee
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  2. Takashi Hibino
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  4. Alejandro M. Viale
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  5. Teruhiro Takabe
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Lee, B.H., Hibino, T., Jo, J. et al. Isolation and characterization of a dnaK genomic locus in a halotolerant cyanobacterium Aphanothece halophytica. Plant Mol Biol 35, 763–775 (1997). https://doi.org/10.1023/A:1005867420619

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