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Open Access Research article

Immobility, inheritance and plasticity of shape of the yeast nucleus

Thomas Hattier12, Erik D Andrulis13 and Alan M Tartakoff12*

Author Affiliations

1 Cell Biology Program, Case Western Reserve University, 10700 Euclid Avenue, Cleveland, OH, 44106 USA

2 Department of Pathology Case Western Reserve University, 10700 Euclid Avenue, Cleveland, OH, 44106, USA

3 Department of Molecular Biology and Microbiology, Case Western Reserve University, 10700 Euclid Avenue, Cleveland, OH, 44106, USA

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BMC Cell Biology 2007, 8:47  doi:10.1186/1471-2121-8-47

Published: 9 November 2007

Additional files

Additional file 1:

Impact of excess Mlp1p. Cells expressing the ER/NE membrane protein, Sec61p-GFP (ATY3255), or Htb2p-mRFP (ATY3256) were induced to overexpress Mlp1p for 5 hr by addition of galactose. Note the conventional distribution of GFP signal at the NE and throughout the peripheral ER (left panel) as well as the roughly spherical chromatin mass (right panel).

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Additional file 2:

Impact of excess Prm3p. Htb2p-mRFP-expressing cells were induced to overexpress GFP-Prm3p for 5 hr by transfer to methionine-free medium (ATY3244). Note the appearance of GFP-positive ring-like structures and a fin at the margin of the nucleus.

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Additional file 3:

Impact of excess Heh1. Htb2p-mRFP-expressing cells were induced galactose to overexpress Heh1p-GFP for 5 hr by addition of 2% (ATY3245). The red and green images have been separated for clarity. Note the massive change of organization of both chromatin and the GFP signal, which often encircles the chromatin.

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Additional file 4:

Impact of excess Heh2p. Htb2p-mRFP-expressing cells were induced to overexpress Heh2p-GFP for 5 hr by addition of 2% galactose (ATY3246). The red and green images have been separated for clarity. Note the massive change of organization of both chromatin and the GFP signal, which generally encircles the chromatin.

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Additional file 5:

Spatial relation of escapades to the nucleolus. Cells expressing the nucleolar marker, Sik1p-mRFP, were induced to express GFP-Esc1p for 5 hr by addition of 2% galactose (ATY3258). Systematic examination of through-focal series detects association of escapades and Sik1p-mRFP in > 90% of cells which have escapades. Nevertheless, in a given section the extent of association often appears only modest.

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Additional file 6:

Spatial relation of escapades to the spindle pole body. Overview comparison of the localization of GFP-tagged Esc1p and the spindle pole body, in a strain (ATY3276) which expresses Spc42p-mRFP and has been induced by addition of 2% galactose for 5 hr. Systematic examination of through-focal series detects association in ~10% of cells which have escapades.

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Additional file 7:

Spatial relation of escapades to a centromere. Overview comparison of the localization of GFP-tagged Esc1p and a centromere, in a strain (ATY2098) which expresses a GFP-lac repressor fusion, an insertion of lac operator arrays near CENIV and Nup49p-GFP. It has been induced by addition of 2% galactose for 5 hr. Systematic examination of through-focal series detects association in ~10% of cells which have escapades.

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Additional file 8:

Spatial relation of escapades to a telomere. Overview comparison of the localization of GFP-tagged escapades and a telomere, in a strain (ATY2097) which expresses a GFP-lac repressor fusion, an insertion of lac operator arrays near telomere XIVL and Nup49p-GFP. It has been induced by addition of 2% galactose for 5 hr. Systematic examination of through-focal series detects association in ~10% of cells which have escapades.

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Additional file 9:

Spatial relation of escapades to Rap1p. Overview localization of Rap 1-GFP in a strain (ATY3275) which expresses Htb2p-mRFP and allows galactose induction of untagged Esc1p. It was induced by addition of 2% galactose for 5 hr. Note that the labeled foci often are at the periphery of the chromatin mass, but – unlike escapades – do not extend centrifugally toward the cytoplasm.

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Additional file 10:

Induction of escapades in cells treated with mating factor. GFP-Esc1p was induced by addition of 2% galactose for 5 hr in cells which had already been treated with 5 μg/ml α-factor for 2 hr (ATY2101). Note that the appearance of escapades is comparable to that illustrated in Figure 1A.

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Additional file 11:

Induction of escapades in cells treated with hydroxyurea. GFP-Esc1p was induced by addition of 2% galactose for 5 hr in cells which had already been treated with 0.1 M hydroxyurea for 2 hr (ATY2101). Note that the appearance of escapades is comparable to that illustrated in Figure 1A.

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Additional file 12:

Induction of escapades in cells treated with nocodazole. GFP-Esc1p was induced by addition of 2% galactose for 5 hr in cells which had already been treated with 15 μg/ml nocodazole 2 hr (ATY2101). Note that the appearance of escapades is comparable to that illustrated in Figure 1A.

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Additional file 13:

Induction of escapades in cells treated with latrunculin A. GFP-Esc1p was induced by addition of 2% galactose for 5 hr in cells which had already been treated with 200 μg/ml latrunculin A 2 hr (ATY2101). Note that the appearance of escapades is comparable to that illustrated in Figure 1A.

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Additional file 14:

Nuclear shape in the absence of Esc1p. The contour of the NE was defined in cells which lack Esc1p by monitoring the distribution of Sec63p-GFP in a corresponding deletion strain (ATY3278).

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