Document Detail


Cell cycle markers for live cell analyses.
MedLine Citation:
PMID:  15701967     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Many cellular processes are regulated by cell cycle dependent changes in protein dynamics and localization. Studying these changes in vivo requires methods to distinguish the different cell cycle stages. Here we demonstrate the use of DNA Ligase I fused to DsRed1 as an in situ marker to identify S phase and the subsequent transition to G2 in live cells. Using this marker, we observed changes in the nuclear distribution of Dnmt1 during cell cycle progression. Based on the different nuclear distribution of DNA Ligase I and Dnmt1 in G2 and G1, we demonstrate that the combination of both proteins allows the direct discrimination of all cell cycle phases using either immunostainings or fusions with fluorescent proteins. These markers are new tools to directly study cell cycle dependent processes in both, fixed and living cells.
Authors:
Hariharan P Easwaran; Heinrich Leonhardt; M Cristina Cardoso
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2005-03-07
Journal Detail:
Title:  Cell cycle (Georgetown, Tex.)     Volume:  4     ISSN:  1551-4005     ISO Abbreviation:  Cell Cycle     Publication Date:  2005 Mar 
Date Detail:
Created Date:  2005-05-06     Completed Date:  2006-05-05     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  101137841     Medline TA:  Cell Cycle     Country:  United States    
Other Details:
Languages:  eng     Pagination:  453-5     Citation Subset:  IM    
Affiliation:
Max Delbrück Center for Molecular Medicine, Berlin, Germany.
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MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Western
Bromodeoxyuridine / pharmacology
COS Cells
Cell Cycle*
Cell Cycle Proteins
Cells, Cultured
Cercopithecus aethiops
DNA (Cytosine-5-)-Methyltransferase / genetics
DNA Ligases / chemistry*
DNA Replication
Electrophoresis, Polyacrylamide Gel
Fluorescent Dyes / chemistry
G2 Phase
Humans
Mice
Microscopy, Fluorescence
Recombinant Fusion Proteins / chemistry
S Phase
Transfection
Chemical
Reg. No./Substance:
0/Cell Cycle Proteins; 0/Fluorescent Dyes; 0/Recombinant Fusion Proteins; 59-14-3/Bromodeoxyuridine; EC 2.1.1.37/DNA (Cytosine-5-)-Methyltransferase; EC 2.1.1.37/DNA (cytosine-5-)-methyltransferase 1; EC 6.5.1.-/DNA Ligases; EC 6.5.1.1/DNA ligase (ATP)

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


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