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Cloning and characterization of two cDNAs encoding casein kinase II catalytic subunits in Arabidopsis thaliana

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Abstract

Two cDNA clones, ATCKA1 and ATCKA2, encoding casein kinase II (CKII) catalytic subunits, were cloned from Arabidopsis thaliana and their nucleotide sequences were determined. Both cDNAs contain 999 bp open reading frames and are 94% identical on the amino acid sequence level. The deduced amino acid sequences of ATCKA1 and ATCKA2 are very similar to that of the human CKII catalytic α subunit (72% homology). Northern blot analysis indicates that the ATCKA1 and ATCKA2 mRNAs are present in all plant organs, but that ATCKA1 transcript levels are quite low compared to those of ATCKA2. Genomic Southern blot analysis suggests that there are at least three CKII genes in the A. thaliana genome. We expressed the ATCKA1 and ATCKA2 cDNAs in Escherichia coli using a pET vector derivative and analyzed the expressed protein in vitro. The expressed ATCKA1 protein phosphorylated casein using either ATP or GTP. This activity was inhibited by heparin, indicating that the expressed protein has activity similar to those reported for animal and yeast CKII.

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Authors and Affiliations

  1. Laboratory of Plant Molecular Biology, Tsukuba Life Science Center, RIKEN (The Institute of Physical and Chemical Research), 3-1-1, Koyadai, 305, Tsukuba, Ibaraki, Japan

    Tsuyoshi Mizoguchi, Kazuko Yamaguchi-Shinozaki, Nobuaki Hayashida & Kazuo Shinozaki

  2. Institute of Biological Sciences, The University of Tsukuba, Tennohdai, 305, Tsukuba, Ibaraki, Japan

    Tsuyoshi Mizoguchi & Hiroshi Kamada

Authors
  1. Tsuyoshi Mizoguchi
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  2. Kazuko Yamaguchi-Shinozaki
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  3. Nobuaki Hayashida
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  4. Hiroshi Kamada
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  5. Kazuo Shinozaki
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Mizoguchi, T., Yamaguchi-Shinozaki, K., Hayashida, N. et al. Cloning and characterization of two cDNAs encoding casein kinase II catalytic subunits in Arabidopsis thaliana . Plant Mol Biol 21, 279–289 (1993). https://doi.org/10.1007/BF00019944

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  • DOI: https://doi.org/10.1007/BF00019944

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