|
|
Relationship between Large-scale Chromatin Organization and Nucleolus in Human Cells
Kalmárová, Markéta ; Raška, Ivan (advisor) ; Nedvídek, Josef (referee) ; Smetana, Karel (referee)
In human cells ribosomal genes are organized as clusters called Nucleolus Organizer Regions (NORs) that are situated on the short arms of acrocentric chromosomes. It was found that essential components of the RNA polymerase I transcription machinery, including Upstream Binding Factor (UBF), can be detected on some NORs, termed "competent" NORs, during mitosis. The competent NORs are believed to be transcriptionally active during interphase. But since individual NORs cannot be observed in the cell nucleus, their interphase status remained unclear. To address this problem, we detected the competent NORs by two commonly used methods, UBF immunofluorescence and silver staining, and combined them with FISH for visualization of rDNA and/or specific chromosomes. We found that the numbers of competent NORs on specific chromosomes were largely conserved in the subsequent cell cycles, with certain NOR-bearing homologues displaying a very stable pattern of competence. Importantly, those and only those NORs, which were loaded with UBF, incorporated bromo-uridine in metaphase after stimulation with roscovitine and in telophase, suggesting that competent and only competent NORs contain ribosomal genes transcriptionally active during interphase. Applying premature chromosome condensation with calyculin A, we visualized...
|
guest ::
