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Synchronization of HeLa cell cultures by inhibition of DNA polymerase alpha with aphidicolin.
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MedLine Citation:
PMID:  6775308     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Both the inhibitory effect of aphidicolin on the replicative alpha-polymerase and the reversibility of its action in vivo (Pedrali-Noy & Spadari, 1979, Biochem. Biophys. Res. Commun. 88, 1194-2002) allow the synchronization of cells in culture. Aphidicolin prevents G1 cells from entering the DNA synthetic period, blocks cells in "S" phase, allows G2, M and G1 cells to continue the cell cycle and to accumulate at the G1/S border. Aphidicolin is a more useful reagent than hydroxyurea and thymidine because it does not affect cell viability or "S" phase duration and does not interfere with the synthesis of dNTPs or DNA polymerases. In fact cells exposed to the drug continue to synthesize all three DNA polymerases alpha, beta and gamma as well as all dNTPs which, when the block is removed, are present at levels optimal for DNA initiation and replication. The technique is simple and can be applied to cells growing in suspension or monolayers and allows one to harvest large quantities of synchronized cells.
Authors:
G Pedrali-Noy; S Spadari; A Miller-Faurès; A O Miller; J Kruppa; G Koch
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Nucleic acids research     Volume:  8     ISSN:  0305-1048     ISO Abbreviation:  Nucleic Acids Res.     Publication Date:  1980 Jan 
Date Detail:
Created Date:  1980-12-18     Completed Date:  1980-12-18     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  0411011     Medline TA:  Nucleic Acids Res     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  377-87     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Aphidicolin
Cell Cycle / drug effects
DNA Polymerase I / metabolism
DNA Polymerase II / antagonists & inhibitors*
DNA Polymerase III / metabolism
DNA-Directed DNA Polymerase / antagonists & inhibitors*
Diterpenes / pharmacology*
Hela Cells / drug effects,  enzymology
Humans
Kinetics
Chemical
Reg. No./Substance:
0/Diterpenes; 38966-21-1/Aphidicolin; EC 2.7.7.-/DNA Polymerase I; EC 2.7.7.-/DNA Polymerase II; EC 2.7.7.-/DNA Polymerase III; EC 2.7.7.7/DNA-Directed DNA Polymerase
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Full Text
Journal Information
Journal ID (nlm-ta): Nucleic Acids Res
ISSN: 0305-1048
ISSN: 1362-4962
Article Information
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Print publication date: Day: 25 Month: 1 Year: 1980
Volume: 8 Issue: 2
First Page: 377 Last Page: 387
ID: 327273
PubMed Id: 6775308

Synchronization of HeLa cell cultures by inhibition of DNA polymerase alpha with aphidicolin.
G Pedrali-Noy
S Spadari
A Miller-Faur?s
A O Miller
J Kruppa
G Koch



Article Categories:
  • Research Article


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