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To study the behavior of newly synthesized RNA molecules during unperturbed mitosis and after induction of aneuploid nuclei, a novel method based on the incorporation of 5-ethynyl uridine was applied. This approach allows revealing transcription in cytological specimens by omitting immunocytochemical procedures. The obtained results indicate that RNAs synthesized in the preceding cell cycle behave similarly as nucleolar proteins and participate in post-mitotic nucleolar assembly not only during unperturbed nuclear divisions. Colocalization of nucleolar proteins and RNAs seems also to occur in aneuploid nuclei devoid of functional nucleolar organizer regions (NORs) or NOR per se. This paper also shows the presence of nucleoplasmic gene expression in aberrant aneuploid nuclei and micronuclei, despite occurrence of their genetic material disorganization. Moreover, experiments showing the effect of transcription and replication inhibition on the induction of aneuploid nuclei were also performed. Presented data shed new light on nucleologenesis and co-existence of nucleolar proteins with RNAs in prenucleolar bodies formed in aneuploid nuclei devoid of functional NORs.
Regardless of the DNA replication stress induced by low concentration of hydroxyurea (HU), root apical meristem cells of Allium cepa keep growing, and some of them override the DNA damage checkpoint mechanisms initiating either premature or an abnormal mitotic chromosome condensation. Prolonged incubation of onion seedlings with HU results in an increased level of immunodetectable proteins sharing epitopes with SUN2, one of the highly conserved elements linking nuclear envelope (NE) to the cytoand nucleoskeletal structures. In addition toNE, phragmoplast and cell plate, our observations extend an array of subcellular compartments at which SUN2-like proteins (SUN2-LPs) are localized. These include cortical preprophase band of microtubules, centromeric regions of ana- and telophase chromosomes, and nuclear bodies (SUN2-NBs) polarly localized in interphase nuclei according to Rabl’s configuration. SUN2- NBs (distinct from fibrillarin-rich Cajal bodies) colocalize with late-replicating areas of heterochromatin and are thought to represent clustered centromeres. Three-dimensional spatial analysis of SUN2-NBs suggests their connections with NE. An enhanced expression and additional localization sites of SUN2-LPs may be correlated with a considerable reprogramming of cellular functions triggered in response to prolonged HU treatment.
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