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Cooperation of two distinct cis-acting elements is necessary for the S phase-specific activation of the wheat histone H3 promoter

Authors

  • Norihiro Ohtsubo,

    1. Department of Botany, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-01, Japan
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    • Present address: Department of Molecular Biology, National Institute of Agrobiological Resources, Tsukuba, Ibaraki 305, Japan.

  • Takuya Nakayama,

    1. Department of Botany, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-01, Japan
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    • Present address: Department of Cell and Developmental Biology, Oregon Health Sciences University, Portland, OR 97201-3098, USA.

  • Hidetaka Kaya,

    1. Department of Botany, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-01, Japan
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  • Rie Terada,

    1. Plantech Research Institute, 1000 Kamoshida, Midori-ku, Yokohama 227, Japan
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    • §

      Present address: National Institute for Basic Biology, 38 Nishigonaka Myodaijicho, Okazaki 444, Japan.

  • Ko Shimamoto,

    1. Plantech Research Institute, 1000 Kamoshida, Midori-ku, Yokohama 227, Japan
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    • Present address: Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-01, Japan.

  • Tetsuo Meshi,

    1. Department of Botany, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-01, Japan
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  • Masaki Iwabuchi

    Corresponding author
    1. Department of Botany, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-01, Japan
      *For correspondence (fax +81 75 753 4142).
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*For correspondence (fax +81 75 753 4142).

Abstract

We have previously shown that the proximal promoter region (−185 to +57) of the wheat histone H3 gene (TH012) is sufficient for regulating S phase-specific expression of a reporter GUS gene. To define the cis-acting element(s) responsible for S phase-specific expression, GUS fusion genes under the control of wild-type or variously mutated H3 promoters were stably introduced into cultured rice Oc cells and their temporal expression was analyzed during the cell cycle by quantitative S1 analysis. The S phase-specific expression of the full-sized promoter (−1716 to +52) was significantly impaired by short internal deletions disrupting the type I element from −175 to −158 (CCACGTCACCaATCCGCG), composed of the Hex (CCACG-TCA) and reverse-oriented Oct (GATCCGCG) motifs. Moreover, the H3 proximal promoters (−184 to +52) harboring base-substitution mutations in either or both of the Hex and Oct motifs could no longer activate gene expression during the S phase. These results indicate that the type I element is the first cis-acting element identified responsible for the S phase-specific expression of plant histone genes. Results also suggested the presence of a redundant cis-acting element(s) responsible for S phase-specific expression in the H3 far-upstream region (−1716 to −185).

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