A novel ataxia-telangiectasia mutated autoregulatory feedback mechanism in murine embryonic stem cells

Robert G. Clyde, Ashley L. Craig, Lucas de Breed, James L. Bown, Leslie Forrester, Borivoj Vojtesek, Graeme Smith, Ted R. Hupp, John W. Crawford

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Abstract

Ataxia-telangiectasia mutated (ATM) is known to play a central role in effecting the DNA damage response that protects somatic cells from potentially harmful mutations, and in this role it is a key anti-cancer agent. However, it also promotes repair of therapeutic damage (e.g. radiotherapy) and so frustrates the efficacy of some treatments. A better understanding of the mechanisms of ATM regulation is therefore important both in prevention and treatment of disease. While progress has been made in elucidating the key signal transduction pathways that mediate damage response in somatic cells, relatively little is known about whether these function similarly in pluripotent embryonic stem (ES) cells where ATM is also implicated in our understanding of adult stem cell ageing and in improvements in regenerative medicine. There is some evidence that different mechanisms may operate in ES cells and that our understanding of the mechanisms of ATM regulation is therefore incomplete. We investigated the behaviour of the damage response signalling pathway in mouse ES cells. We subjected the cells to the DNA-damaging agent doxorubicin, a drug that induces double-strand breaks, and measured ATM expression levels. We found that basal ATM gene expression was unaffected by doxorubicin treatment. However, following ATM kinase inhibition using a specific ATM inhibitor, we observed a significant increase in ATM and ataxia-telangiectasia and Rad3 related transcription. We demonstrate the use of a dynamical modelling approach to show that these results cannot be explained in terms of known mechanisms. Furthermore, we show that the modelling approach can be used to identify a novel feedback process that may underlie the anomalies in the data. The predictions of the model are consistent both with our in vitro experiments and with in vivo studies of ATM expression in somatic cells in mice, and we hypothesize that this feedback operates in both somatic and ES cells in vivo. The results point to a possible new target for ATM inhibition that overcomes the restorative potential of the proposed feedback.
Original languageEnglish
Pages (from-to)1167-1177
Number of pages11
JournalJournal of the Royal Society Interface
Volume6
Issue number41
DOIs
StatePublished - 6 Dec 2009

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Status Epilepticus
Ataxia Telangiectasia
Embryonic Stem Cells
Stem cells
Feedback
Inborn Errors Amino Acid Metabolism
DNA
Common Bile Duct Diseases
Supravalvular Aortic Stenosis
Cellophane
Anthralin
Signal transduction
Regenerative Medicine
Transcription
Gene expression
Repair
Oxidoreductases Acting on CH-NH Group Donors
Doxorubicin
Inhibition (Psychology)
Radiotherapy

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Clyde, R. G., Craig, A. L., Breed, L. D., Bown, J. L., Forrester, L., Vojtesek, B., ... Crawford, J. W. (2009). A novel ataxia-telangiectasia mutated autoregulatory feedback mechanism in murine embryonic stem cells. Journal of the Royal Society Interface, 6(41), 1167-1177. DOI: 10.1098/rsif.2008.0538

Clyde, Robert G.; Craig, Ashley L.; Breed, Lucas de; Bown, James L.; Forrester, Leslie; Vojtesek, Borivoj; Smith, Graeme; Hupp, Ted R.; Crawford, John W. / A novel ataxia-telangiectasia mutated autoregulatory feedback mechanism in murine embryonic stem cells.

In: Journal of the Royal Society Interface, Vol. 6, No. 41, 06.12.2009, p. 1167-1177.

Research output: Contribution to journalArticle

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Clyde, RG, Craig, AL, Breed, LD, Bown, JL, Forrester, L, Vojtesek, B, Smith, G, Hupp, TR & Crawford, JW 2009, 'A novel ataxia-telangiectasia mutated autoregulatory feedback mechanism in murine embryonic stem cells' Journal of the Royal Society Interface, vol 6, no. 41, pp. 1167-1177. DOI: 10.1098/rsif.2008.0538

A novel ataxia-telangiectasia mutated autoregulatory feedback mechanism in murine embryonic stem cells. / Clyde, Robert G.; Craig, Ashley L.; Breed, Lucas de; Bown, James L.; Forrester, Leslie; Vojtesek, Borivoj; Smith, Graeme; Hupp, Ted R.; Crawford, John W.

In: Journal of the Royal Society Interface, Vol. 6, No. 41, 06.12.2009, p. 1167-1177.

Research output: Contribution to journalArticle

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AU - Clyde,Robert G.

AU - Craig,Ashley L.

AU - Breed,Lucas de

AU - Bown,James L.

AU - Forrester,Leslie

AU - Vojtesek,Borivoj

AU - Smith,Graeme

AU - Hupp,Ted R.

AU - Crawford,John W.

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Clyde RG, Craig AL, Breed LD, Bown JL, Forrester L, Vojtesek B et al. A novel ataxia-telangiectasia mutated autoregulatory feedback mechanism in murine embryonic stem cells. Journal of the Royal Society Interface. 2009 Dec 6;6(41):1167-1177. Available from, DOI: 10.1098/rsif.2008.0538