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. 2004 Sep 7;101(36):13221-6.
doi: 10.1073/pnas.0402958101. Epub 2004 Aug 26.

Nuclear body movement is determined by chromatin accessibility and dynamics

Sabine M Görisch  1 , Malte Wachsmuth, Carina Ittrich, Christian P Bacher, Karsten Rippe, Peter Lichter
Affiliations

Nuclear body movement is determined by chromatin accessibility and dynamics

Sabine M Görisch et al. Proc Natl Acad Sci U S A. .

Abstract

Promyelocytic leukemia (PML) and Cajal bodies are mobile subnuclear organelles, which are involved in activities like RNA processing, transcriptional regulation, and antiviral defense. A key parameter in understanding their biological functions is their mobility. The diffusion properties of PML and Cajal bodies were compared with a biochemically inactive body formed by aggregates of murine Mx1 by using single-particle tracking methods. The artificial Mx1-yellow fluorescent protein body showed a very similar mobility compared with PML and Cajal bodies. The data are described quantitatively by a mechanism of nuclear body movement consisting of two components: diffusion of the body within a chromatin corral and its translocation resulting from chromatin diffusion. This finding suggests that the body mobility reflects the dynamics and accessibility of the chromatin environment, which might target bodies to specific nuclear subcompartments where they exert their biological function.

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Figures

Fig. 1.
Fig. 1.
Mx1-YFP colocalizes with nuclear bodies. Confocal images of Mx1-YFP (A), Mx1-YFP with PML bodies (B), and Mx1-YFP with Cajal bodies (C) are shown. Mx1-YFP is shown in green, PML and Cajal bodies in red, and 4′,6-diamidino-2-phenylindole staining in blue. (D) Electron micrograph showing a Mx1-YFP body. (Scale bars: 10 μm, A–C; 1 μm, D.)
Fig. 2.
Fig. 2.
In vivo images. Confocal sections of Mx1-YFP (A), PML bodies (B), and Cajal bodies (C) are shown. The nuclear shape is outlined by a dotted line. (Scale bars: 10 μm.)
Fig. 3.
Fig. 3.
A single nuclear body displays different types of motion over time. Confined, obstructed, and free diffusion as well as directed motion for individual Mx1-YFP (A1–A4), PML (B1–B4), and Cajal bodies (C1–C4) are shown. The respective α values for the classification are noted.
Fig. 4.
Fig. 4.
Single-particle tracking analysis of nuclear bodies. Shown are average MSD plots for all Mx1-YFP (A), PML bodies (B), and Cajal bodies (C) examined. The fit for obstructed diffusion is shown in red, and the fit for diffusion of particles in corrals is shown in blue, both with weighted errors. The χ2 values for the red and the blue fits are 0.34 and 0.27 for Mx1-YFP, 0.59 and 0.31 for PML bodies, and 0.47 and 0.28 for Cajal bodies. Average log-log plots for Mx1-YFP (D), PML bodies (E), and Cajal bodies (F) are shown. The normal log-log plots are shown in red with the α values 0.34, 0.27, and 0.33 for Mx1-YFP, PML, and Cajal bodies, respectively. The log-log plots with exclusion of Db are shown in blue. The fitis a horizontal line obtained from the mean value of the curve points. The α values are accordingly 1.
Fig. 5.
Fig. 5.
Schematic illustration of nuclear body diffusion. The nuclear body is enclosed in a chromatin corral, defined by the chromatin accessibility in which they diffuse with Db as demonstrated by the deformation of the chromatin. The chromatin corrals translocate within the nucleus with the Dc indicated by the red arrow.
Fig. 6.
Fig. 6.
Analysis of single nuclear bodies. For all bodies studied the rc, Dc, and Db values according to Eq. 3 were calculated individually. The resulting distribution of rc, Dc, and Db is plotted for Mx1-YFP (A1–A3), PML (B1–B3), and Cajal bodies (C1–C3). All histograms show essentially a distribution with a single peak indicative of only one species of each nuclear body type.
Fig. 7.
Fig. 7.
Single-particle tracking analysis of dense chromatin regions. (A) Mx1-YFP particles in a cell line stably expressing H2A-CFP (SW13H2A-CFP). An example for an evaluated dense chromatin block is labeled by *. (Scale bar: 10 μm.) (B) Average MSD plot for 15 evaluated regions. The fit for obstructed diffusion is shown in red (χ2 = 0.032), and the fit for diffusion of particles in corrals is shown in blue (χ2 = 0.018).
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