Document Detail


Macroscopic folding and replication of the homogeneously staining region in late S phase leads to the appearance of replication bands in mitotic chromosomes.
MedLine Citation:
PMID:  15454579     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The chromosomal G/R bands are alternating domains differing in their nucleotide sequence biases. The bands are also related to the time of replication: pulse-labeling during S phase makes the replication sites as visible as replication bands that are close to the G/R bands in mitotic chromosomes. We previously showed that a plasmid bearing a mammalian replication origin efficiently generated a chromosomal homogeneously staining region (HSR). Here, we analyze the replication of this artificial HSR and show that it was replicated at the last stage of S phase. The HSR was composed of plasmid repeats only; nonetheless, we found that replication sites pulse-labeled during late S phase appeared as bands in the mitotic HSR and their number was dependent on the length of the HSR. Therefore, replication bands might not arise from sequence information per se. To understand the chronological order of appearance of replication sites, we performed a double pulse-chase experiment using IdU and CldU. Replication of the entire HSR required 100-120 minutes. During this period, the replicated sites appeared as bands at the first and last stages, but in between were apparently scattered along the entire HSR. An analysis of S-phase nuclei revealed that the replication started at the periphery of the globular HSR domain, followed by initiation in the internal domain. The replicated HSR appeared as a ring or a pair of extended spirals in late G2-phase nuclei. To account for these findings, we present a model in which the HSR is folded as a coiled-coil structure that is replicated from the outside to the inside in S phase nuclei.
Authors:
Noriaki Shimizu; Kenta Shingaki
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2004-09-28
Journal Detail:
Title:  Journal of cell science     Volume:  117     ISSN:  0021-9533     ISO Abbreviation:  J. Cell. Sci.     Publication Date:  2004 Oct 
Date Detail:
Created Date:  2004-10-14     Completed Date:  2005-05-16     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0052457     Medline TA:  J Cell Sci     Country:  England    
Other Details:
Languages:  eng     Pagination:  5303-12     Citation Subset:  IM    
Affiliation:
Faculty of Integrated Arts and Sciences, Hiroshima University, 1-7-1 Kagamiyama, Higashi-hiroshima, 739-8521, Japan. shimizu@hiroshima-u.ac.jp
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MeSH Terms
Descriptor/Qualifier:
Cell Line, Tumor
Cell Nucleus / metabolism
Chromosomes / ultrastructure*
Coloring Agents / pharmacology
G2 Phase
Heterochromatin / ultrastructure
Humans
In Situ Hybridization, Fluorescence
Microscopy, Confocal
Mitosis
Nucleic Acid Conformation
Nucleic Acid Hybridization
Plasmids / metabolism
S Phase
Thymidine / metabolism
Time Factors
Transfection
Chemical
Reg. No./Substance:
0/Coloring Agents; 0/Heterochromatin; 50-89-5/Thymidine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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