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Publication Detail

Title: ATRMec1 phosphorylation-independent activation of Chk1 in vivo.

Authors: Chen, Yinhuai; Caldwell, Julie M; Pereira, Elizabeth; Baker, Robert W; Sanchez, Yolanda

Published In J Biol Chem, (2009 Jan 02)

Abstract: The conserved protein kinase Chk1 is a player in the defense against DNA damage and replication blocks. The current model is that after DNA damage or replication blocks, ATR(Mec1) phosphorylates Chk1 on the non-catalytic C-terminal domain. However, the mechanism of activation of Chk1 and the function of the Chk1 C terminus in vivo remains largely unknown. In this study we used an in vivo assay to examine the role of the C terminus of Chk1 in the response to DNA damage and replication blocks. The conserved ATR(Mec1) phosphorylation sites were essential for the checkpoint response to DNA damage and replication blocks in vivo; that is, that mutation of the sites caused lethality when DNA replication was stalled by hydroxyurea. Despite this, loss of the ATR(Mec1) phosphorylation sites did not change the kinase activity of Chk1 in vitro. Furthermore, a single amino acid substitution at an invariant leucine in a conserved domain of the non-catalytic C terminus restored viability to cells expressing the ATR(Mec1) phosphorylation site-mutated protein and relieved the requirement of an upstream mediator for Chk1 activation. Our findings show that a single amino acid substitution in the C terminus, which could lead to an allosteric change in Chk1, allows it to bypass the requirement of the conserved ATR(Mec1) phosphorylation sites for checkpoint function.

PubMed ID: 18984588 Exiting the NIEHS site

MeSH Terms: Amino Acid Substitution; Checkpoint Kinase 1; DNA Damage/drug effects; DNA Damage/physiology*; DNA Replication/drug effects; DNA Replication/physiology*; Enzyme Activation/physiology; Hydroxyurea/pharmacology; MAP Kinase Kinase 1/genetics; MAP Kinase Kinase 1/metabolism*; Nucleic Acid Synthesis Inhibitors/pharmacology; Phosphorylation/drug effects; Phosphorylation/physiology; Point Mutation; Protein Kinases/genetics; Protein Kinases/metabolism*; Protein Structure, Tertiary/physiology; Saccharomyces cerevisiae Proteins/genetics; Saccharomyces cerevisiae Proteins/metabolism*; Saccharomyces cerevisiae/enzymology*; Saccharomyces cerevisiae/genetics

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