Microtubule Organisation in Dictyostelium

Abstract

Dictyostelium amoebae contain a radial array of microtubules emanating from a single microtubule‐organising centre called centrosome that is bound to the cytosolic face of the nucleus. Their centrosome contains no centrioles but consists of a layered core surrounded by a corona harbouring microtubule nucleation centres. It duplicates in prophase of a closed mitosis and organises a central spindle that drives centrosome separation and chromosome segregation. Dictyostelium microtubules exhibit a differential dynamic behaviour during interphase. Growth and shrinkage is observed only in the periphery but not in the pericentrosomal region. During mitosis, when centrosomes possess no corona, microtubules behave quite dynamically in formation of a central spindle and astral microtubules. Microtubules are associated with a couple of conserved proteins (MAPs), which are involved in centrosome biogenesis and the cross talk of microtubule tips with the actin cell cortex. The latter becomes evident in cytokinesis, when centrosomes with their attached microtubules participate in the positioning of cleavage furrows.

Key Concepts

  • Dictyostelium amoebae contain a nucleus‐associated centrosome that serves as the only microtubule‐organising centre.
  • The Dictyostelium centrosome contains no centrioles, but consists of a three‐layered core structure surrounded by a microtubule‐nucleating corona.
  • If compared to the three major plaques of the yeast spindle pole body, the entire core structure of the Dictyostelium centrosome appears equivalent to the central plaque, while the corona plays a similar role as the inner and outer plaques.
  • Dictyostelium centrosomes duplicate at the onset of mitosis.
  • Dictyostelium amoebae show a closed type of mitosis with a persisting nuclear envelope.
  • Dictyostelium microtubules are quite dynamic during mitosis but show growth and shrinkage only in the periphery during interphase.
  • Microtubule plus ends regulate actin dynamics at the cell cortex.
  • Dictyostelium amoebae are a useful model to study the role of the centrosome and microtubules in cell dynamics and disease.

Keywords: Dictyostelium; centrosome; MAP; microtubules; mitosis; cytokinesis

Figure 1. The Dictyostelium centrosome cycle. Nuclei, chromosomes and centrosomes with attached microtubules are shown in schematic cross sections of different cell cycle stages, except for prometaphase where a view to the nuclear surface is depicted. The corresponding immunofluorescence microscopy images show staining of the centrosomes with anti‐CP224 (red), the nuclear envelope with the centrosome/nucleus linkage with anti‐Sun1 (green) and DNA with DAPI (blue). The detailed process is described in the text.
Figure 2. Live cell imaging of cell expressing GFP‐EB1 reveals microtubule interactions with the cell cortex and positioning of cleavage furrows with respect to microtubule arrays. The sequence shows late mitosis of a cell with two supernumerary centrosomes, which are not associated with a central spindle and are involved in formation of a cytoplast during cytokinesis. Selected time points of a movie shown. Cells were viewed under agar overlay. Each image represents a brightest point z‐projection of five confocal slices with a distance of 1 µm each (Perkin Elmer Ultraview, 1.3/100×, 12‐bit CCD at 2 × 2 binning, frame rate 1.4 fr s−1, time lapse 10 s)
Figure 3. FRAP experiments of cells expressing GFP‐α‐tubulin reveal differential dynamics of microtubules in the periphery (a) and the pericentrosomal area (b). While fluorescence recovers almost completely at peripheral microtubules, there is almost no recovery in the pericentrosomal region despite of the centrosome itself (see text for discussion). Cells were viewed under agar overlay. Each image represents a brightest point z‐projection of five confocal slices with a distance of 0.5 µm each (Zeiss LSM710, 1.4/63× lens, pinhole 2 AU, frame rate 3 fr s−1). Bar 4 µm. Matthias Samereier and Ralph Gräf, University of Potsdam, unpublished results.
Figure 4. Live cell imaging of cells expressing GFP‐α‐tubulin reveals decomposition of the microtubule system in prophase (150 s) and re‐growth of a radial microtubule array in telophase (1080 s). Selected time points of a movie starting at the G2/M transition are shown. Cells were viewed under agar overlay. Each image represents a brightest point z‐projection of five confocal slices with a distance of 1.6 µm each (Zeiss LSM510, 512 × 512 pixel, 1.4/63× lens, pinhole 2 AU, frame rate 2 fr s−1, time lapse 10 s)
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Further reading

Artemenko Y, Lampert TJ and Devreotes PN (2014) Moving towards a paradigm: common mechanisms of chemotactic signaling in Dictyostelium and mammalian leukocytes. Cellular and Molecular Life Sciences 71: 3711–3747.

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White EA and Glotzer M (2012) Centralspindlin: at the heart of cytokinesis. Cytoskeleton 69: 882–892.

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How to Cite close
Gräf, Ralph(Feb 2015) Microtubule Organisation in Dictyostelium. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0021852.pub2]