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DNA replication and chromosome positioning throughout the interphase in three dimensional space of plant nuclei

Němečková Alžběta, Veronika Koláčková, Vrána Jan, Doležel Jaroslav, Hřibová Eva
doi: https://doi.org/10.1101/2020.04.02.021857
Němečková Alžběta
Institute of Experimental Botany of the Czech Academy of Sciences, Centre of the Region Hana for Biotechnological and Agricultural Research, Olomouc, Czech Republic
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Veronika Koláčková
Institute of Experimental Botany of the Czech Academy of Sciences, Centre of the Region Hana for Biotechnological and Agricultural Research, Olomouc, Czech Republic
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Vrána Jan
Institute of Experimental Botany of the Czech Academy of Sciences, Centre of the Region Hana for Biotechnological and Agricultural Research, Olomouc, Czech Republic
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Doležel Jaroslav
Institute of Experimental Botany of the Czech Academy of Sciences, Centre of the Region Hana for Biotechnological and Agricultural Research, Olomouc, Czech Republic
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Hřibová Eva
Institute of Experimental Botany of the Czech Academy of Sciences, Centre of the Region Hana for Biotechnological and Agricultural Research, Olomouc, Czech Republic
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  • For correspondence: hribova@ueb.cas.cz
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Abstract

Despite the recent progress, our understanding of the principles of plant genome organization and its dynamics in three-dimensional space of interphase nuclei remains limited. In this study, DNA replication timing and interphase chromosome positioning was analyzed in seven Poaceae species differing in genome size. A multidisciplinary approach combining newly replicated DNA labelling by EdU, nuclei sorting by flow cytometry, three-dimensional immuno-FISH, and confocal microscopy revealed similar replication timing order for telomeres and centromeres as well as for euchromatin and heterochromatin in all seven species. The Rabl configuration of chromosomes that lay parallel to each other and their centromeres and telomeres are localized at opposite nuclear poles, was observed in wheat, oat, rye and barley with large genomes, as well as in Brachypodium with a small genome. On the other hand, chromosomes of rice with a small genome and maize with relatively large genome did not assume proper Rabl configuration. In all species, the interphase chromosome positioning inferred from the location of centromeres and telomeres was stable throughout the interphase. These observations extend earlier studies indicating a more complex relation between genome size and interphase chromosome positioning, which is controlled by factors currently not known.

Highlight Telomere and centromere replication timing and interphase chromosome positioning in seven grass species differing in genome size indicates a more complex relation between genome size and the chromosome positioning.

Footnotes

  • E-mail addresses: Němečková A. - nemeckova{at}ueb.cas.cz, Koláčková V. - kolackova{at}ueb.cas.cz, Vrána J. - jan.vrana{at}osu.cz, Doležel J. - dolezel{at}ueb.cas.cz

  • Abbreviations

    3D
    three-dimensional
    CenH3
    centromere-specific variant of histone H3
    BrdU
    5-bromo-2’-deoxyuridine
    EdU
    5-ethynyl-2’-deoxyuridine
    FISH
    fluorescence in situ hybridization
    PAA
    polyacrylamide
    rDNA
    ribosomal deoxyribonucleic acid
    ROI
    region of interest
    RT
    room temperature
  • Copyright 
    The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
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    Posted April 02, 2020.
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    DNA replication and chromosome positioning throughout the interphase in three dimensional space of plant nuclei
    Němečková Alžběta, Veronika Koláčková, Vrána Jan, Doležel Jaroslav, Hřibová Eva
    bioRxiv 2020.04.02.021857; doi: https://doi.org/10.1101/2020.04.02.021857
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    DNA replication and chromosome positioning throughout the interphase in three dimensional space of plant nuclei
    Němečková Alžběta, Veronika Koláčková, Vrána Jan, Doležel Jaroslav, Hřibová Eva
    bioRxiv 2020.04.02.021857; doi: https://doi.org/10.1101/2020.04.02.021857

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