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First published online June 28, 2004
doi: 10.1242/10.1242/jcs.01203

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S. pombe meiotic linear elements contain proteins related to synaptonemal complex components

¹ Institute of Botany, University of Vienna, Rennweg 14, A-1030 Vienna, Austria
² NWCRF Cancer Research Unit, School of Biological Sciences, University of Wales Bangor, Deiniol Road, Bangor, Gwynedd, LL57 2UW, UK
³ Bioinformatics Group, Research Institute of Molecular Pathology (IMP), Dr. Bohr-Gasse 7, A-1030 Vienna, Austria

View larger version (70K): [in a new window]   Fig. 1. (a) Multiple sequence alignment of the homologous C-termini of Red1-related proteins in S. cerevisiae (Sc), Kluyveromyces lactis (Kl), Candida glabrata (Cg) and S. pombe (Sp). Sequences are indicated with the species, followed by a database accession number [databases used are Genpept (Sc, Kl, Sp) and Génolevures (Cg) (Feldmann, 2000)]. Sequences are labelled with a four-level conservation shading using similarity groups. (b) Multiple sequence alignment of the HORMA domain (Aravind and Koonin, 1998) from Hop1-related proteins of Sc, Debaryomyces hansenii (Dh), Giardia intestinalis (Gi), Arabidopsis thaliana (At), Caenorhabditis elegans (Ce), Mus musculus (Mm) and Sp. Sequences are indicated with the species, followed by a database accession number [databases used are Génolevures (Dh) (Feldmann, 2000), Pompep (Sp) and Genpept (all remaining sequences)]. Sequences are labelled with a four-level conservation shading using similarity groups.

View larger version (98K): [in a new window]   Fig. 2. LEs as observed after silver staining in the electron microscope (EM) (top) and by immunostaining of Rec10 (bottom). The classification of LEs in the EM is according to Bähler et al. (Bähler et al., 1993). Nuclei containing many (up to >50) Rec10-po sitive spots (designated here as class Ia) appear inconspicuous in the EM. The empty nucleus in the EM (top left, class assignment not applicable) might correspond by its shape and size to immunostained class Ia. Classes I (Ib) and IIa are identified by both immunostaining and EM as containing individual threads and meshes of threads, respectively. Class IIb appears in the EM as bundles of LEs that probably correspond to the long, thick Rec10-positive structures. Silver-stained individual LEs of heterogeneous lengths, which prevail during later stages of meiosis (7-10 hours), were designated as class III (Bähler et al., 1993). The corresponding Rec10-LEs are classified as Ib. A class-Ib nucleus from a late time point in meiosis (8 hours in sporulation medium) is shown for comparison. Scale bar, 5 µm.

View larger version (30K): [in a new window]   Fig. 3. Proportions of different LE classes in nuclei of the wild type at different times in sporulation medium. A typical time course is shown. For the classification of LEs, see Fig. 2. 200 nuclei were evaluated for each time point.

View larger version (54K): [in a new window]   Fig. 4. Examples of Rec10-positive LEs (red) in (a) the wild type (WT) and various mutants. Two typical nuclei are shown for each strain. (b) In the rec10-155 mutant, LEs are completely missing, whereas, in the rec10-109 mutant (c), LEs develop into a few dots or short lines. (d) In a rec8 strain, Rec10 is organized into thick threads resembling class IIb LEs. (e) In the rec12-152 mutant strain, all classes of LEs occur (a class IIa and a class IIb nucleus are shown). (f) In a mek1 strain, Rec10 assembles into wild-type-like LEs, whereby classes Ia and Ib are more abundant than the others. Chromatin is stained blue with DAPI. Scale bar, 5 µm.

View larger version (61K): [in a new window]   Fig. 5. Hop1 and Mek1 localization to Rec10 LEs shown by immunostaining of Rec10, Hop1 and Mek1-HA. Regions where Hop1 or Mek1 (green) colocalize with Rec10 (red) appear orange in the merge image. Chromatin is stained blue with DAPI. The images show Hop1 localization to: (a) class Ib LEs of the wild type (WT); (b) residual LEs in the rec10-109 mutant; (c) LEs in a rec12-152 mutant strain; (d) aberrant LEs in the rec8 mutant; and (e) LEs in a mek1 strain. Some short LEs are lacking Hop1 and Hop1 localization to longer LEs is discontinuous. Mek1 forms dots on short LEs in class I nuclei (f) but covers most parts of LE bundles in (rare) class IIb nuclei (g). Scale bar, 5 µm.

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