RT Journal Article
SR Electronic
T1 DNA replication and chromosome positioning throughout the interphase in three dimensional space of plant nuclei
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.04.02.021857
DO 10.1101/2020.04.02.021857
A1 Němečková Alžběta
A1 Veronika Koláčková
A1 Vrána Jan
A1 Doležel Jaroslav
A1 Hřibová Eva
YR 2020
UL http://biorxiv.org/content/early/2020/04/02/2020.04.02.021857.abstract
AB 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.3Dthree-dimensionalCenH3centromere-specific variant of histone H3BrdU5-bromo-2’-deoxyuridineEdU5-ethynyl-2’-deoxyuridineFISHfluorescence in situ hybridizationPAApolyacrylamiderDNAribosomal deoxyribonucleic acidROIregion of interestRTroom temperature