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Spatio‐temporal dynamics at rDNA foci: Global switching between DNA replication and transcription
Author(s) -
Pliss Artem,
Koberna Karel,
Večeřová Jaromíra,
Malínský Jan,
Mašata Martin,
Fialová Markéta,
Raška Ivan,
Berezney Ronald
Publication year - 2004
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.20317
Subject(s) - fibrillarin , biology , nucleolus , transcription (linguistics) , ribosomal dna , chromatin , rna polymerase i , microbiology and biotechnology , gene , rna , genetics , rna polymerase , linguistics , philosophy , phylogenetics , cytoplasm
We have investigated the in situ organization of ribosomal gene (rDNA) transcription and replication in HeLa cells. Fluorescence in situ hybridization (FISH) revealed numerous rDNA foci in the nucleolus. Each rDNA focus corresponds to a higher order chromatin domain containing multiple ribosomal genes. Multi‐channel labeling experiments indicated that, in the majority of cells, all the rDNA foci were active in transcription as demonstrated by co‐localization with signals to transcription and fibrillarin, a protein involved in ribosomal RNA processing. In some cells, however, a small portion of the rDNA foci did not overlap with signals to transcription and fibrillarin. Labeling for DNA replication revealed that those rDNA foci inactive in transcription were restricted to the S‐phase of the cell cycle and were replicated predominantly from mid to late S‐phase. Electron microscopic analysis localized the nucleolar transcription, replication, and fibrillarin signals to the dense fibrillar components of the nucleolus and at the borders of the fibrillar centers. We propose that the rDNA foci are the functional units for coordinating replication and transcription of the rRNA genes in space and time. This involves a global switching mechanism, active from mid to late S‐phase, for turning off transcription and turning on replication at individual rDNA foci. Once all the rRNA genes at individual foci are replicated, these higher order chromatin domains are reprogrammed for transcription. © 2004 Wiley‐Liss, Inc.