Document Detail

Centrosome replication in hydroxyurea-arrested CHO cells expressing GFP-tagged centrin2.
MedLine Citation:
PMID:  17606999     Owner:  NLM     Status:  MEDLINE    
Centrosome duplication is tightly coupled with the cell cycle and neither too many nor too few centrosomes are induced in a normal cell. To study how centrosome assembly is regulated, we analyzed the abnormal process of multiple centrosome replications in Chinese hamster ovary (CHO) cells induced by hydroxyurea (HU), which is known to uncouple the centrosome cycle from the cell cycle. Green fluorescent protein (GFP)-tagged centrin2 expressed in CHO cells labels both centrioles and the pericentriolar material (PCM). Counting fluorescent spots of GFP-centrin in synchronized cells showed that in G(1)/S-arrested cells, centrioles are initially duplicated in a template manner. Further treatment with HU overrides the suppression of excess centriole/centrosome replication in a cell where the full complement of centrioles/centrosomes already exists. Time-lapse fluorescence microscopy revealed that small centrin-containing foci emerged in the cytoplasm during HU treatment. These foci are surrounded by a PCM cloud and their number continuously increases as cells are exposed to HU for longer periods of time. Both the centrosome and cytoplasmic foci are highly mobile, continuously changing their position in a manner dependent on microtubules/microtubule dynamics. The centrosome number increases as small foci grow in size and resolve into recognizable centrosomes. As this occurs in a random fashion, the cells arrested longer with HU induced highly heterogeneous numbers of centrosomes.
Ryoko Kuriyama; Yasuhiko Terada; Kyung S Lee; Christopher L C Wang
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, N.I.H., Intramural    
Journal Detail:
Title:  Journal of cell science     Volume:  120     ISSN:  0021-9533     ISO Abbreviation:  J. Cell. Sci.     Publication Date:  2007 Jul 
Date Detail:
Created Date:  2007-07-03     Completed Date:  2007-11-28     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0052457     Medline TA:  J Cell Sci     Country:  England    
Other Details:
Languages:  eng     Pagination:  2444-53     Citation Subset:  IM    
Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, MN 55455, USA.
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MeSH Terms
CHO Cells
Calcium-Binding Proteins / genetics,  metabolism*
Cell Cycle Proteins / genetics,  metabolism*
Centrioles / metabolism*
Centrosome / metabolism*
Green Fluorescent Proteins / metabolism
Hydroxyurea / pharmacology
Mitotic Spindle Apparatus / metabolism
Recombinant Fusion Proteins / metabolism
Grant Support
Reg. No./Substance:
0/CETN2 protein, human; 0/Calcium-Binding Proteins; 0/Cell Cycle Proteins; 0/Recombinant Fusion Proteins; 127-07-1/Hydroxyurea; 147336-22-9/Green Fluorescent Proteins

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