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Originally published In Press as doi:10.1074/jbc.M101756200 on March 26, 2001

J. Biol. Chem., Vol. 276, Issue 25, 22772-22778, June 22, 2001
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Two Phases of Chromatin Decondensation during Dedifferentiation of Plant Cells
DISTINCTION BETWEEN COMPETENCE FOR CELL FATE SWITCH AND A COMMITMENT FOR S PHASE*

Jing Zhao, Nadya Morozova, Leor Williams, Laurence Libs, Yigal Avivi, and Gideon GrafiDagger

From the Department of Plant Sciences, The Weizmann Institute of Science, Rehovot 76100, Israel

Cellular dedifferentiation is the major process underlying totipotency, regeneration, and formation of new stem cell lineages in multicellular organisms. In animals it is often associated with carcinogenesis. Here, we used tobacco protoplasts (plant cells devoid of cell wall) to study changes in chromatin structure in the course of dedifferentiation of mesophyll cells. Using flow cytometry and micrococcal nuclease analyses, we identified two phases of chromatin decondensation prior to entry of cells into S phase. The first phase takes place in the course of protoplast isolation, following treatment with cell wall degrading enzymes, whereas the second occurs only after protoplasts are induced with phytohormones to re-enter the cell cycle. In the absence of hormonal application, protoplasts undergo cycles of chromatin condensation/decondensation and die. The ubiquitin proteolytic system was found indispensable for protoplast progression into S phase, being required for the second but not the first phase of chromatin decondensation. The emerging model suggests that cellular dedifferentiation proceeds by two functionally distinct phases of chromatin decondensation: the first is a transitory phase that confers competence for cell fate switch, which is followed, under appropriate conditions, by a second proteasome-dependent phase representing a commitment for the mitotic cycle. These findings might have implications for a wide range of dedifferentiation-driven cellular processes in higher eukaryotes.

* This research was supported by a grant from the Israel Science Foundation (ISF).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Plant Sciences, The Weizmann Inst. of Science, Rehovot 76100, Israel. Tel.: 972-8-934-3505; Fax: 972-8-934-4181; E-mail: gideon.grafi@weizmann.ac.il.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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