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

0538924 - ÚEB 2021 RIV GB eng J - Journal Article
Němečková, Alžběta - Koláčková, Veronika - Vrána, Jan - Doležel, Jaroslav - Hřibová, Eva
DNA replication and chromosome positioning throughout the interphase in three-dimensional space of plant nuclei.
Journal of Experimental Botany. Roč. 71, č. 20 (2020), s. 6262-6272. ISSN 0022-0957. E-ISSN 1460-2431
R&D Projects: GA MŠMT(CZ) LM2015062; GA ČR(CZ) GA17-14048S; GA MŠMT(CZ) EF16_019/0000827
Institutional support: RVO:61389030
Keywords : genome-wide analysis * s-phase * arabidopsis-thaliana * chromatin packing * timing program * allium-cepa * organization * telomeres * domains * dynamics * DNA replication * EdU labeling * flow cytometry * Poaceae * Rabl configuration * S phase * three-dimensional fluorescence in situ hybridization (3D-FISH)
OECD category: Biochemistry and molecular biology
Impact factor: 6.992, year: 2020
Method of publishing: Open access
http://doi.org/10.1093/jxb/eraa370

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, three-dimensional 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.
Permanent Link: http://hdl.handle.net/11104/0316663