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Transcriptional activation of tobacco E2F is repressed by co-transfection with the retinoblastoma-related protein: cyclin D expression overcomes this repressor activity

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Abstract

Evidence is emerging that the E2F family of transcription factors plays an important role in the regulation of gene expression at the G1/S transition in plants. Here, we show that in the tobacco proliferating cell nuclear antigen (PCNA), whose transcript is specifically expressed at G1/S phase, the two E2F binding sites are synergistically responsible for transcriptional activation at G1/S phase in synchronized tobacco BY-2 cells transformed with promoter constructs fused to a reporter gene. In addition, we have isolated the tobacco DP cDNA (NtDP) and showed that significant activation of the reporter gene was observed in transient expression assays by concomitantly transfecting with plasmids expressing NtE2F and NtDP. This transcriptional activation was repressed by co-transfection with a plasmid expressing NtRBR1; in vitro pull-down assays also revealed that NtRBR1 binds directly to NtE2F, thereby potentially blocking the transcriptional activation of NtE2F. Importantly, this repressor activity was cancelled when NtRBR1 was further co-transfected with a plasmid expressing cyclin D but not with cyclin A or cyclin B. These results are discussed with respect to the repression activity of NtRBR1 on the NtE2F/NtDP complex.

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Abbreviations

CDK:

cyclin-dependent serine/threonine protein kinase

Rb:

retinoblastoma

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Authors and Affiliations

  1. Graduate School of Biological Sciences, Nara Institute of Science and Technology (NAIST), Takayama 8916-5, Ikoma, Nara, 630-0101, Japan

    Kenji Uemukai, Hidekazu Iwakawa, Ko Kato, Atsuhiko Shinmyo & Masami Sekine

  2. Institute of Advanced Biosciences, Keio University, Tsuruoka, Yamagata, 997-0035, Japan

    Shunichi Kosugi

  3. Institute of Biotechnology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QT, United Kingdom

    Sarah de Uemukai & James AH Murray

  4. Institut des Sciences Végétales, CNRS, Avenue de la Terrasse, 91198, Gif sur Yvette, France

    Eva Kondorosi

  5. Department of Regulation of Biological Signals, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya, 464-8601, Japan

    Masaki Ito

Authors
  1. Kenji Uemukai
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  2. Hidekazu Iwakawa
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  3. Shunichi Kosugi
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  4. Sarah de Uemukai
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  5. Ko Kato
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  6. Eva Kondorosi
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  7. James AH Murray
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  8. Masaki Ito
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  9. Atsuhiko Shinmyo
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  10. Masami Sekine
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Correspondence to Masami Sekine.

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Uemukai, K., Iwakawa, H., Kosugi, S. et al. Transcriptional activation of tobacco E2F is repressed by co-transfection with the retinoblastoma-related protein: cyclin D expression overcomes this repressor activity. Plant Mol Biol 57, 83–100 (2005). https://doi.org/10.1007/s11103-004-6601-x

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  • DOI: https://doi.org/10.1007/s11103-004-6601-x

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