Čeština
EnglishDNA replication and chromosome positioning throughout the interphase in three-dimensional space of plant nuclei
Němečková A., Koláčková V., Vrána J., Doležel J., Hřibová E.
JOURNAL OF EXPERIMENTAL BOTANY 71: 6262-6272, 2020
Klíčová slova: DNA replication, EdU labeling, flow cytometry, Poaceae, Rabl configuration, S phase, three-dimensional fluorescence in situ hybridization (3D-FISH)
Abstrakt: Despite much recent progress, our understanding of the principles of plant genome organization and its dynamics in three-dimensional space of interphase nuclei remains surprisingly limited. Notably, it is not clear how these processes could be affected by the size of a plant’s nuclear genome. In this study, DNA replication timing and interphase chromosome positioning were analyzed in seven Poaceae species that differ in their genome size. To provide a comprehensive picture, a suite of advanced, complementary methods was used: labeling of newly replicated DNA by ethynyl-2'-deoxyuridine, isolation of nuclei at particular cell cycle phases by flow cytometric sorting, threedimensional immunofluorescence in situ hybridization, and confocal microscopy. Our results revealed conserved dynamics of DNA replication in all species, and a similar replication timing order for telomeres and centromeres, as well as for euchromatin and heterochromatin regions, irrespective of genome size. Moreover, stable chromosome positioning was observed while transitioning through different stages of interphase. These findings expand upon earlier studies in suggesting that a more complex interplay exists between genome size, organization of repetitive DNA sequences along chromosomes, and higher order chromatin structure and its maintenance in interphase, albeit controlled by currently unknown factors.
DOI: 10.1093/jxb/eraa370
Fulltext: kontaktujte autory z ÚEB
Autoři z ÚEB: Alžběta Doležalová, Jaroslav Doležel, Eva Hřibová, Veronika Koláčková, Jan Vrána
JOURNAL OF EXPERIMENTAL BOTANY 71: 6262-6272, 2020
Klíčová slova: DNA replication, EdU labeling, flow cytometry, Poaceae, Rabl configuration, S phase, three-dimensional fluorescence in situ hybridization (3D-FISH)
Abstrakt: Despite much recent progress, our understanding of the principles of plant genome organization and its dynamics in three-dimensional space of interphase nuclei remains surprisingly limited. Notably, it is not clear how these processes could be affected by the size of a plant’s nuclear genome. In this study, DNA replication timing and interphase chromosome positioning were analyzed in seven Poaceae species that differ in their genome size. To provide a comprehensive picture, a suite of advanced, complementary methods was used: labeling of newly replicated DNA by ethynyl-2'-deoxyuridine, isolation of nuclei at particular cell cycle phases by flow cytometric sorting, threedimensional immunofluorescence in situ hybridization, and confocal microscopy. Our results revealed conserved dynamics of DNA replication in all species, and a similar replication timing order for telomeres and centromeres, as well as for euchromatin and heterochromatin regions, irrespective of genome size. Moreover, stable chromosome positioning was observed while transitioning through different stages of interphase. These findings expand upon earlier studies in suggesting that a more complex interplay exists between genome size, organization of repetitive DNA sequences along chromosomes, and higher order chromatin structure and its maintenance in interphase, albeit controlled by currently unknown factors.
DOI: 10.1093/jxb/eraa370
Fulltext: kontaktujte autory z ÚEB
Autoři z ÚEB: Alžběta Doležalová, Jaroslav Doležel, Eva Hřibová, Veronika Koláčková, Jan Vrána