Functional uncoupling of MCM helicase and DNA polymerase activities activates the ATR-dependent checkpoint

The ATR-dependent DNA damage response pathway can respond to a diverse group of lesions as well as inhibitors of DNA replication. Using the Xenopus egg extract system, we show that lesions induced by UV irradiation and cis-platinum cause the functional uncoupling of MCM helicase and DNA polymerase a...

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Vydané v:Genes & development Ročník 19; číslo 9; s. 1040
Hlavní autori: Byun, Tony S, Pacek, Marcin, Yee, Muh-ching, Walter, Johannes C, Cimprich, Karlene A
Médium: Journal Article
Jazyk:English
Vydavateľské údaje: United States 01.05.2005
Predmet:
Adaptor Proteins, Signal Transducing - metabolism
Animals
Aphidicolin - metabolism
Aphidicolin - pharmacology
Ataxia Telangiectasia Mutated Proteins
Cell Cycle Proteins - metabolism
Cell-Free System
Checkpoint Kinase 1
Chromatin - metabolism
DNA Damage
DNA Helicases - metabolism
DNA Primers
DNA Replication - physiology
DNA, Single-Stranded - drug effects
DNA, Single-Stranded - metabolism
DNA-Directed DNA Polymerase - metabolism
Phosphorylation
Plasmids - genetics
Protein Kinases - metabolism
Protein-Serine-Threonine Kinases - metabolism
Xenopus
Xenopus Proteins - metabolism
ISSN:0890-9369
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Shrnutí:The ATR-dependent DNA damage response pathway can respond to a diverse group of lesions as well as inhibitors of DNA replication. Using the Xenopus egg extract system, we show that lesions induced by UV irradiation and cis-platinum cause the functional uncoupling of MCM helicase and DNA polymerase activities, an event previously shown for aphidicolin. Inhibition of uncoupling during elongation with inhibitors of MCM7 or Cdc45, a putative helicase cofactor, results in abrogation of Chk1 phosphorylation, indicating that uncoupling is necessary for activation of the checkpoint. However, uncoupling is not sufficient for checkpoint activation, and DNA synthesis by Polalpha is also required. Finally, using plasmids of varying size, we demonstrate that all of the unwound DNA generated at a stalled replication fork can contribute to the level of Chk1 phosphorylation, suggesting that uncoupling amplifies checkpoint signaling at each individual replication fork. Taken together, these observations indicate that functional uncoupling of MCM helicase and DNA polymerase activities occurs in response to multiple forms of DNA damage and that there is a general mechanism for generation of the checkpoint-activating signal following DNA damage.
Bibliografia:ObjectType-Article-2
SourceType-Scholarly Journals-1
ObjectType-Feature-1
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ISSN:0890-9369
DOI:10.1101/gad.1301205