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Persistent mechanical linkage between sister chromatids throughout anaphase

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Abstract

In budding yeast, we have found that sister rDNA arrays marked with fluorescent probes can be visualized as two distinguishable strands during metaphase. Upon anaphase, these arm loci are drawn into the spindle, where they adopt a cruciform-like structure and stretch 2.5-fold as they migrate to the poles. Therefore, while sister rDNA arrays appear separated in metaphase, mechanical linkages between sister arm loci persist throughout anaphase in yeast, as shown in grasshopper spermatocytes (Paliulis and Nicklas 2004). These linkages are partially dependent on the protector of cohesin, SGO1. In anaphase, the spatially regulated dissolution of these mechanical linkages serves to prevent premature sister separation and restrain the rate of spindle elongation. Thus, sister separation is temporally controlled and linkages between sister chromatids contribute to the regulation of anaphase spindle elongation.

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Acknowledgments

We thank Irem Unlu and Sena Özşeker for their hard work imaging rDNA-imbedded lacO arrays. We thank Jeff Molk, Julian Haase, Elaine Yeh, Gidi Shemer, Dr. Doug Koshland, and members of the Bloom laboratory for advice, assistance, and critical readings of the manuscript. This work was funded by the National Institute of Health RO1 Grant GM32238 (K. Bloom).

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Authors and Affiliations

  1. Department of Biology, University of North Carolina at Chapel Hill, CB# 3280, Coker Hall, Chapel Hill, NC, 27599-3280, USA

    Benjamin D. Harrison & Kerry Bloom

  2. Department of Embryology, Carnegie Institution, Baltimore, MD, 21218, USA

    Margaret L. Hoang

Authors
  1. Benjamin D. Harrison
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  2. Margaret L. Hoang
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  3. Kerry Bloom
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Corresponding author

Correspondence to Kerry Bloom.

Additional information

Communicated by F. Uhlmann

Electronic supplementary materials

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Supplementary Figure 1

rDNA segregation trajectory in mating cells tracked using CDC14-GFP (rDNA) and SPC29-RFP (spindle poles). a Trans-image of zygote. b Metaphase with CDC14 in green and SPC29 in red. c Anaphase. Scale bar in a represents 2 μm. (GIF 157 kb)

High resolution image (TIFF 6412 kb)

Supplementary Figure 2

rDNA segregation trajectory in mating cells tracked using Net1-GFP. a Trans-image of zygote. b Metaphase with NET1 in green and SPC29 in red. c Anaphase. Scale bar in a represents 2 μm. (GIF 62 kb)

High resolution image (TIFF 1962 kb)

Supplementary Figure 3

Tracking a single strand of Net1 fluorescence through mitosis. A single strand of Net1-GFP was marked by bleaching a section. b The strand prebleach and the arrow head in c points to the bleached region. After anaphase, the bleached region is found on one strand in the daughter cell (d, arrow) and not on either strand in the mother cell (n = 8). The scale bar in a represents 2(m. (GIF 122 kb)

High resolution image (TIFF 3713 kb)

Supplementary Table 1

Fluorescent strand dimensions in the first metaphase after mating using NET1-GFP. Values are not statistically different than values using CDC14-GFP. (DOC 29 kb)

Supplementary Table 2

Fluorescent strand dimension in the first anaphase after mating using NET1-GFP. Values are not statistically different than values using CDC14-GFP. (DOC 30 kb)

Supplementary Movie 1

Metaphase and anaphase of a SGO1 zygote. Images in the movie were taken at 1 min intervals. The bright foci in the middle of the zygote represent the kinetochore clusters (NUF2-GFP). The fluorescent signal above and below the NUF2-GFP foci are the rDNA arrays marked with CDC14-GFP. (AVI 14388 kb)

Supplementary Movie 2

Metaphase and anaphase of a sgo1Δ zygote. Images in the movie were taken at 1 min intervals. Red foci represent the spindle poles marked by SPC29-RFP. The green fluorescence is the rDNA array marked with CDC14-GFP. (AVI 26772 kb)

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Harrison, B.D., Hoang, M.L. & Bloom, K. Persistent mechanical linkage between sister chromatids throughout anaphase. Chromosoma 118, 633–645 (2009). https://doi.org/10.1007/s00412-009-0224-6

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  • DOI: https://doi.org/10.1007/s00412-009-0224-6

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