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

Hall-Jackson, Clare A; Cross, Darren A.E.; Morrice, Nick; Smythe, Carl

doi:10.1038/sj.onc.1203077

Cited by 208 publications

(159 citation statements)

References 55 publications

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“…18 To test if ATM/ATR pathways control the inhibition of Plk1 phosphorylation at T210 after treatment with adriamycin, we pretreated HEK293-T cells with caffeine, an ATM/ATR inhibitor, in the presence of adriamycin and nocodazole. [26][27][28] Pretreatment with 5 mM caffeine partially spared the phosphorylation of T210 (Fig. 4, lower panel, lane 5) and decreased Plk1 inhibition after adriamycin treatment (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…4, two middle panels, lanes 5 and 7). At these concentrations, caffeine inhibits both ATM and ATR; [26][27][28] and UCN-01 inhibits both Chk1 and Chk2. 31 Equal extents of restoration of T210 phosphorylation and Plk1 activity by treatment with caffeine and UCN-01 are consistent with the fact that ATM/ATR and Chk1/Chk2 act in the same signaling pathways.…”

mentioning

confidence: 99%

“…17 Since ATM/ATR pathways inhibit Plk1 activity in response to DNA damage, we used caffeine, an inhibitor of ATM and ATR, to test if they mediate the inhibition of Plk1 phosphorylation. [26][27][28] Involvement of ATM/ATR-regulated kinases, Chk2 and Chk1, was assessed by treatment with UCN-01, an inhibitor of Chk1 and Chk2 kinases. Initial reports describing the effects of UCN-01 on recombinant Chk1 and Chk2 kinases found that Chk1 is much more sensitive to UCN-01 (IC 50 = 11 nM) than is Chk2 (IC 50 = 1040 nM).…”

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confidence: 99%

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Phosphorylation of Plk1 at S137 and T210 is Inhibited in Response to DNA Damage

Tsvetkov

Stern²

2004

Cell Cycle

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Section: Resultsmentioning

confidence: 99%

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Phosphorylation of Plk1 at S137 and T210 is Inhibited in Response to DNA Damage

Tsvetkov

Stern²

2004

Cell Cycle

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“…It is well known that agents that uncouple the ATM/ATR-CHK1/CHK2 axis can ablate the G 2 DNA damage checkpoint. These include inhibitors of ATM/ATR (for example, caffeine) (Blasina et al, 1999b;Hall-Jackson et al, 1999) and CHK1 (for example, UCN-01). Attenuation of the checkpoint induces unscheduled activation of CDK1 and mitotic catastrophe (Chan et al, 1999;Castedo et al, 2004a, b;Vogel et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Generation of an indestructible cyclin B1 by caspase-6-dependent cleavage during mitotic catastrophe

2008

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Overriding the G 2 DNA damage checkpoint permits precocious entry into mitosis that ultimately leads to mitotic catastrophe. Mitotic catastrophe is manifested by an unscheduled activation of CDK1, caspase activation and apoptotic cell death. We found that although cyclin B1 was required for mitotic catastrophe, it was cleaved into a B35 kDa protein by a caspase-dependent mechanism during the process. Cyclin B1 cleavage occurred after Asp123 in the motif ILVD 123 k, and mutation of this motif attenuated the cleavage. Cleavage was abolished by a pan-caspase inhibitor as well as by specific inhibitors for the effector caspase-6 and the initiator caspase-8. Cleavage created a truncated cyclin B1 lacking part of the NH 2 -terminal regulatory domain that included the destruction box sequence. Although cleavage of cyclin B1 itself was not absolutely required for mitotic catastrophe, expression of the truncated product enhanced cell death. In support of this, ectopic expression of this truncated cyclin B1 was not only sufficient to induce mitotic block and apoptosis but also enhanced mitotic catastrophe induced by ionizing radiation and caffeine. These data underscore a possible linkage between mitotic and apoptotic functions by caspase-dependent processing of mitotic activators.

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“…Caffeine has been demonstrated to override DNA damage checkpoints in a number of organisms by inhibiting the DNA damage checkpoint transducer ATR (Homo sapiens)/Mec1 (S. cerevisiae)/Rad3 (S. pombe) (Schlegel and Pardee, 1986;Osman and McCready, 1998;Hall-Jackson et al, 1999;Moser et al, 2000;Vaze et al, 2002;Liberi et al, 2005). Therefore, we compared the effects of caffeine addition to a culture overexpressing TOP3 Y356F after 3 h of MMS treatment.…”

Section: Top3 Functions In Recombinational Repairmentioning

confidence: 99%

Top3 Processes Recombination Intermediates and Modulates Checkpoint Activity after DNA Damage

Mankouri

Hickson

2006

MBoC

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Mutation of TOP3 in Saccharomyces cerevisiae causes poor growth, hyperrecombination, and a failure to fully activate DNA damage checkpoints in S phase. Here, we report that overexpression of a dominant-negative allele of TOP3, TOP3 Y356F , which lacks the catalytic (decatenation) activity of Top3, causes impaired S-phase progression and the persistence of abnormal DNA structures (X-shaped DNA molecules) after exposure to methylmethanesulfonate. The impaired S-phase progression is due to a persistent checkpoint-mediated cell cycle delay and can be overridden by addition of caffeine. Hence, the catalytic activity of Top3 is not required for DNA damage checkpoint activation, but it is required for normal S-phase progression after DNA damage. We also present evidence that the checkpoint-mediated cell cycle delay and persistence of X-shaped DNA molecules resulting from overexpression of TOP3 Y356F are downstream of Rad51 function. We propose that Top3 functions in S phase to both process homologous recombination intermediates and modulate checkpoint activity. INTRODUCTIONTopoisomerases are highly conserved proteins that catalyze topological rearrangements in the structure of DNA and are required for many aspects of DNA metabolism. Of the three topoisomerases in yeast, the function(s) of the sole type IA topoisomerase, Top3, remains most poorly understood. However, it is likely that Top3 fulfills important roles in vivo, because deletion of TOP3 in Saccharomyces cerevisiae causes hyperrecombination, sensitivity to genotoxic agents, meiotic defects, and poor growth due to accumulation of cells with a late S/G 2 content of DNA (Wallis et al., 1989;Gangloff et al., 1994Gangloff et al., , 1999Chakraverty et al., 2001). Similarly, deletion of top3 ϩ in Schizosaccharomyces pombe causes lethality due to chromosome missegregation and accumulation of DNA double-strand breaks (Goodwin et al., 1999;Maftahi et al., 1999;Oh et al., 2002;Win et al., 2004). Whereas lower eukaryotes generally contain only one type IA topoisomerase, human cells, like most vertebrates, possess at least two Top3 homologues, hTOPIII␣ and hTOPIII␤ (Hanai et al., 1996;Ng et al., 1999). In mice, mutation of TOP3␣ causes embryonic lethality (Li and Wang, 1998), whereas mutation of TOP3␤ causes a shortened life span (Kwan et al., 2003). Interestingly, in S. cerevisiae and S. pombe, deletion of SGS1 or rqh1 ϩ can largely suppress the phenotypes caused by deletion of TOP3 or top3 ϩ , respectively (Gangloff et al., 1994;Goodwin et al., 1999;Maftahi et al., 1999;Chakraverty et al., 2001). Because both SGS1 and rqh1 ϩ belong to the same family of proteins, the RecQ DNA helicases, this intriguing genetic interaction has led to the suggestion that type IA topoisomerases may be functionally associated with RecQ helicases. Indeed, both Sgs1 and Rqh1 physically interact with Top3 in their respective organisms (Gangloff et al., 1994;Bennett et al., 2000;Fricke et al., 2001;Onodera et al., 2002;Laursen et al., 2003;Ahmad and Stewart, 2005;Ui et al., 2005), and a close as...

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

Cited by 208 publications

References 55 publications

Phosphorylation of Plk1 at S137 and T210 is Inhibited in Response to DNA Damage

Phosphorylation of Plk1 at S137 and T210 is Inhibited in Response to DNA Damage

Generation of an indestructible cyclin B1 by caspase-6-dependent cleavage during mitotic catastrophe

Top3 Processes Recombination Intermediates and Modulates Checkpoint Activity after DNA Damage

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