Interphase Cell Cycle Dynamics of a Late-Replicating, Heterochromatic Homogeneously Staining Region: Precise Choreography of Condensation/Decondensation and Nuclear Positioning

doi:10.1083/jcb.140.5.975

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J. Cell Biol., Volume 140, Number 5, March 9, 1998 975-989

Interphase Cell Cycle Dynamics of a Late-Replicating, Heterochromatic Homogeneously Staining Region: Precise Choreography of Condensation/Decondensation and Nuclear Positioning

Gang Li, Gail Sudlow, and Andrew S. Belmont

Department of Cell and Structural Biology, University of Illinois, Urbana-Champaign, Urbana, Illinois 61801

Recently we described a new method for in situ localization of specific DNA sequences, based on lac operator/repressor recognition(Robinett, C.C., A. Straight, G. Li, C. Willhelm, G. Sudlow, A.Murray, and A.S. Belmont. 1996. J. Cell Biol. 135:1685-1700).We have applied this methodology to visualize the cell cycle dynamicsof an ~90 Mbp, late-replicating, heterochromatic homogeneouslystaining region (HSR) in CHO cells, combining immunostaining withdirect in vivo observations. Between anaphase and early G1, theHSR extends approximately twofold to a linear, ~0.3-µm-diam chromatid,and then recondenses to a compact mass adjacent to the nuclearenvelope. No further changes in HSR conformation or position areseen through mid-S phase. However, HSR DNA replication is precededby a decondensation and movement of the HSR into the nuclear interior4-6 h into S phase. During DNA replication the HSR resolves intolinear chromatids and then recondenses into a compact mass; thisis followed by a third extension of the HSR during G2/ prophase.Surprisingly, compaction of the HSR is extremely high at all stagesof interphase. Preliminary ultrastructural analysis of the HSRsuggests at least three levels of large-scale chromatin organizationabove the 30-nm fiber.

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