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Published online 25 February 2003. doi:10.1083/jcb.200211103
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© The Rockefeller University Press, 0021-9525/2003/3/685 $5.00
The Journal of Cell Biology, Volume 160, Number 5, 685-697

Article

 Chromosome order in HeLa cells changes during mitosis and early G1, but is stably maintained during subsequent interphase stages

Joachim Walter, Lothar Schermelleh, Marion Cremer, Satoshi Tashiro and Thomas Cremer

Department of Biology II, Ludwig Maximilians University (LMU), 80333 Munich, Germany

Address correspondence to Thomas Cremer, Dept. Biologie II, Lehrstuhl für Anthropologie und Humangenetik, LMU, Richard-Wagner-Strasse 10/I, 80333 München, Germany. Tel.: 49 (89) 2180-6710. Fax: 49 (89) 2180-6719. E-mail: Thomas.Cremer{at}lrz.uni-muenchen.de

Whether chromosomes maintain their nuclear positions during interphase and from one cell cycle to the next has been controversially discussed. To address this question, we performed long-term live-cell studies using a HeLa cell line with GFP-tagged chromatin. Positional changes of the intensity gravity centers of fluorescently labeled chromosome territories (CTs) on the order of several µm were observed in early G1, suggesting a role of CT mobility in establishing interphase nuclear architecture. Thereafter, the positions were highly constrained within a range of ~1 µm until the end of G2. To analyze possible changes of chromosome arrangements from one cell cycle to the next, nuclei were photobleached in G2 maintaining a contiguous zone of unbleached chromatin at one nuclear pole. This zone was stably preserved until the onset of prophase, whereas the contiguity of unbleached chromosome segments was lost to a variable extent, when the metaphase plate was formed. Accordingly, chromatin patterns observed in daughter nuclei differed significantly from the mother cell nucleus. We conclude that CT arrangements were stably maintained from mid G1 to late G2/early prophase, whereas major changes of CT neighborhoods occurred from one cell cycle to the next. The variability of CT neighborhoods during clonal growth was further confirmed by chromosome painting experiments.

Key Words: nuclear architecture; chromosome territories; chromatin dynamics; cell cycle; photobleaching


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