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The DNA-binding protease, CND41, and the degradation of ribulose-1,5-bisphosphate carboxylase/oxygenase in senescent leaves of tobacco

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Abstract

Plastids bear their own genome, organized into DNA–protein complexes (nucleoids). Recently, we identified a DNA-binding protease (CND41) in the chloroplast nucleoids of cultured tobacco (Nicotiana tabacum L.) cells. In this study, we examine the biochemical function of this novel DNA-binding protease, particularly in senescent leaves, because antisense tobacco with a reduced amount of CND41 showed retarded senescence. Nitrogen-depletion experiments clearly showed that CND41 antisense tobacco maintained green leaves and constant protein levels, especially ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), throughout the whole plant, whereas wild-type tobacco showed marked senescence and the reduction of protein levels in the lower leaves. In vitro analyses confirmed that CND41 showed proteolytic activity at physiological pH when denatured Rubisco was used as the substrate. These results suggest that CND41 is involved in Rubisco degradation and the translocation of nitrogen during senescence. The possible regulation of protease activity of CND41 through DNA-binding is discussed.

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Abbreviations

CABP :

2-Carboxyarabinitol-1,5-bisphosphate

CBB :

Coomassie Brilliant Blue

GS :

Glutamine synthetase

OEC33 :

The extrinsic 33-kDa protein in the oxygen-evolving complex

Rubisco :

Ribulose 1,5-bisphosphate carboxylase/oxygenase

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Acknowledgements

We thank the late Dr. A. Watanabe at the University of Tokyo, Dr. T. Yamaya at Tohoku University, Dr. M. Sugita at Nagoya University and Dr. M. Shigeoka at Kinki University for their kind gifts of anti-OEC33, anti-glutamine synthetase, anti-chloroplast ribonucleoproteins and anti-ascorbate peroxidase antibodies, respectively. This research was supported in part by grants-in-aid from the Ministry of Education, Science, Culture and Sports of Japan (to F.S.; No. 09274103) and the Future Program Grant JSPS-RFTF00L01606 from the Japan Society for the Promotion of Science (to F.S.). Y. Kato and S. Murakami contributed equally to this work.

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Authors and Affiliations

  1. Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, 606-8502, Kyoto, Japan

    Yusuke Kato, Yumiko Yamamoto, Yoshihiko Kondo & Fumihiko Sato

  2. Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, 606-8502, Kyoto, Japan

    Shinya Murakami, Hiroshi Chatani & Fumihiko Sato

  3. The Institute of Physical and Chemical Research (RIKEN), 351-0189, Wako, Japan

    Takeshi Nakano

  4. Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, 630-0101, Nara, Japan

    Akiho Yokota

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  1. Yusuke Kato
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  2. Shinya Murakami
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  3. Yumiko Yamamoto
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Correspondence to Fumihiko Sato.

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Kato, Y., Murakami, S., Yamamoto, Y. et al. The DNA-binding protease, CND41, and the degradation of ribulose-1,5-bisphosphate carboxylase/oxygenase in senescent leaves of tobacco. Planta 220, 97–104 (2004). https://doi.org/10.1007/s00425-004-1328-0

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