Protein kinase mutants of human ATR increase sensitivity to UV and ionizing radiation and abrogate cell cycle checkpoint control

Wright, James R.; Keegan, Kathleen; Herendeen, Daniel R.; Bentley, Nicola J.; Carr, Antony; Hoekstra, Merl F.; Concannon, Patrick

doi:10.1073/pnas.95.13.7445

Cited by 218 publications

(162 citation statements)

References 21 publications

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“…These large proteins lack kinase motifs and activity. Size fractionation studies indicate that both ATM and ATR are also found in large protein complexes of about 1 ± 2 million daltons (Wright et al, 1998;Shalev, unpublished). Similar unpublished ®ndings were mentioned by Chen and Lee (1996) and Gately et al (1998).…”

Section: Dsb Repair and Chromatin Alterationsmentioning

confidence: 99%

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ATM: A mediator of multiple responses to genotoxic stress

1999

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The ATM protein kinase is the product of the gene responsible for the pleiotropic recessive disorder ataxiatelangiectasia. ATM-de®cient cells show enhanced sensitivity and greatly reduced responses to genotoxic agents that generate DNA double strand breaks (DSBs), such as ionizing radiation and radiomimetic chemicals, but exhibit normal responses to DNA adducts and base modi®cations induced by other agents. Therefore, DSBs are most likely the predominant signal for the activation of ATM-mediated pathways. Identi®cation of the ATM gene triggered extensive research aimed at elucidating the numerous functions of its large multifaceted protein product. While ATM has both nuclear and cytoplasmic functions, this review will focus on its roles in the nucleus where it plays a central role in the very early stages of damage detection and serves as a master controller of cellular responses to DSBs. By activating key regulators of multiple signal transduction pathways, ATM mediates the e cient induction of a signaling network responsible for repair of the damage, and for cellular recovery and survival.

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Section: Dsb Repair and Chromatin Alterationsmentioning

confidence: 99%

“…The mammalian family member most closely related to ATM is the ATR/FRP1 protein; like its homologs in yeast (Rad3 of S. pombe and Mec1 of S. cerevisiae), it mediates cellular responses to unreplicated or damaged DNA (Cliby et al, 1998;Wright et al, 1998).…”

Section: The Atm Proteinmentioning

confidence: 99%

ATM: A mediator of multiple responses to genotoxic stress

1999

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“…ATR is structurally related to ATM and can phosphorylate both serine 15 and serine 37 of human p53 in vitro, albeit with 20-fold lower activity as compared with ATM under the same conditions (Canman et al, 1998;Tibbetts et al, 1999). ATR has been proposed to have overlapping function with ATM on the basis that kinase-inactive mutants of ATR cause hypersensitivity to g or x-radiation and hydroxyurea, and abrogate radiation-induced checkpoint control (Cliby et al, 1998;Wright et al, 1998). Analyses with kinase-dead mutants also revealed that ATR can block the maintenance, but not the initiation, of serine 15 phosphorylation in response to IR, suggesting that ATM and ATR act sequentially in the DNA damage response (Tibbetts et al, 1999).…”

Section: Protein Kinase Activities Involved In Phosphorylating P53 Inmentioning

confidence: 99%

Mechanisms of switching on p53: a role for covalent modification?

1999

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The p53 protein plays a pivotal role in activating and integrating adaptive cellular responses to a wide range of environmental stresses. Activation of p53 can occur by di erent molecular routes, depending on the nature of the activating signal. Central to the activation process, by whichever route, is the destabilization of the p53-MDM2 interaction. The molecular mechanisms which activate p53 involve elements of post-translational modi®cation, protein stabilization and protein-protein interaction. Two central themes are emerging from recent work in this area. The ®rst is that there are common events in the p53 activation process among di erent activating pathways. The second is that activation involves not just a single molecular event such as disruption of the p53-MDM2 interaction, but a series of sequential events the nature of which is governed by the type of activating stimulus. This review summarizes our current knowledge of the p53 activation process in response to two stimuli, DNA damage and activated oncogenes, and considers the contribution made by multisite phosphorylation in determining the nature of the p53 response.

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“…More recently, a human homologue of rad3 has been described Cimprich et al, 1996). This protein, termed ATR (for ATM and rad3 related) can complement the UV sensitivity of mec1 mutants when expressed in S. cerevisiae, and expression of kinase-dead ATR protein in human cells causes sensitivity to DNA damaging agents and defects in cell cycle checkpoints (Cliby et al, 1998;Wright et al, 1998). Immunoprecipitates of ATR have associated protein kinase activity which is reduced in immunoprecipitates of putative kinase-dead protein (Canman et al, 1998;Keegan et al, 1996;Sarkaria et al, 1998;Tishko et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

ATR is a caffeine-sensitive, DNA-activated protein kinase with a substrate specificity distinct from DNA-PK

et al. 1999

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ATR is a large, 4300 kDa protein containing a carboxy-terminus kinase domain related to PI-3 kinase, and is homologous to the ATM gene product in human cells and the rad3/MEC1 proteins in yeast. These proteins, together with the DNA-PK, are part of a new family of PI-3 kinase related proteins. All members of this family play important roles in checkpoints which operate to permit cell survival following many forms of DNA damage. We have expressed ATR protein in HEK293 cells and puri®ed the protein to near-homogeneity. We show that pure ATR is a protein kinase which is activated by circular single-stranded, doublestranded or linear DNA. Thus ATR is a new member of a sub-family of PIK related kinases, founded by the DNA-PK, which are activated in the presence of DNA. Unlike DNA-PK, ATR does not appear to require Ku proteins for its activation by DNA. We show directly that, like ATM and DNA-PK, ATR phosphorylates the genome surveillance protein p53 on serine 15, a site which is up-regulated in response to DNA damage. In addition, we ®nd that ATR has a substrate speci®city similar to, but unique from, the DNA-PK in vitro, suggesting that these proteins have overlapping but distinct functions in vivo. Finally, we ®nd that the kinase activity of ATR in the presence and absence of DNA is suppressed by ca eine, a compound which is known to induce loss of checkpoint control. Our results are consistent with the notion that ATR plays a role in monitoring DNA structure and phosphorylation of proteins involved in the DNA damage response pathways.

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Protein kinase mutants of human ATR increase sensitivity to UV and ionizing radiation and abrogate cell cycle checkpoint control

Cited by 218 publications

References 21 publications

ATM: A mediator of multiple responses to genotoxic stress

ATM: A mediator of multiple responses to genotoxic stress

Mechanisms of switching on p53: a role for covalent modification?

ATR is a caffeine-sensitive, DNA-activated protein kinase with a substrate specificity distinct from DNA-PK

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