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The differential loading of two barley CENH3 variants into distinct centromeric substructures is cell type- and development-specific

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Abstract

The organization of centromeric chromatin of diploid barley (Hordeum vulgare) encoding two (α and β) CENH3 variants was analysed by super-resolution microscopy. Antibody staining revealed that both CENH3 variants are organized in distinct but intermingled subdomains in interphase, mitotic and meiotic centromeres. Artificially extended chromatin fibres illustrate that these subdomains are formed by polynucleosome clusters. Thus, a CENH3 variant-specific loading followed by the arrangement into specific intermingling subdomains forming the centromere region appears. The CENH3 composition and transcription vary among different tissues. In young embryos, most interphase centromeres are composed of both CENH3 variants, while in meristematic root cells, a high number of nuclei contain βCENH3 mainly dispersed within the nucleoplasm. A similar distribution and no preferential arrangement of the two CENH3 variants in relationship to the spindle poles suggest that both homologs meet the same function in metaphase cells.

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Abbreviations

CENH3:

Centromeric histone H3 variant

CENP-C:

Centromere protein C

SIM:

Structured illumination microscopy

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Acknowledgments

This work was supported by the German Federal Ministry of Education and Research (FKZ 0315965). We are grateful to Oda Weiß, Katrin Kumke and Karla Meier for excellent technical assistance and to Karin Lipfert for help with artwork.

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Author notes

  1. Ali Mohammad Banaei-Moghaddam

    Present address: Department of Plant Sciences, School of Biology, College of Science, University of Tehran, P.O. Box 14155-6455, Tehran, Iran

Authors and Affiliations

  1. Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben, Corrensstr. 3, 06466, Stadt Seeland, Germany

    Takayoshi Ishii, Raheleh Karimi-Ashtiyani, Ali Mohammad Banaei-Moghaddam, Veit Schubert, Jörg Fuchs & Andreas Houben

Authors
  1. Takayoshi Ishii
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  2. Raheleh Karimi-Ashtiyani
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  3. Ali Mohammad Banaei-Moghaddam
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Corresponding author

Correspondence to Andreas Houben.

Additional information

Responsible Editor: Hans de Jong.

Takayoshi Ishii and Raheleh Karimi-Ashtiyani contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Rotating barley metaphase chromosomes showing the centromeric localization of α and βCENH3. (AVI 4910 kb)

Enlargment of the centromeric region shown in Suppl. Movie 1. (AVI 5970 kb)

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(DOC 117 kb)

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Ishii, T., Karimi-Ashtiyani, R., Banaei-Moghaddam, A.M. et al. The differential loading of two barley CENH3 variants into distinct centromeric substructures is cell type- and development-specific. Chromosome Res 23, 277–284 (2015). https://doi.org/10.1007/s10577-015-9466-8

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  • DOI: https://doi.org/10.1007/s10577-015-9466-8

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