• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. Jan 2, 1998; 17(1): 159–169.
PMCID: PMC1170367

Overexpression of a kinase-inactive ATR protein causes sensitivity to DNA-damaging agents and defects in cell cycle checkpoints.

Abstract

ATR, a phosphatidylinositol kinase-related protein homologous to ataxia telangiectasia mutated (ATM), is important for the survival of human cells following many forms of DNA damage. Expression of a kinase-inactive allele of ATR (ATRkd) in human fibroblasts causes increased sensitivity to ionizing radiation (IR), cis-platinum and methyl methanesulfonate, but only slight UV radiation sensitivity. ATRkd overexpression abrogates the G2/M arrest after exposure to IR, and overexpression of wild-type ATR complements the radioresistant DNA synthesis phenotype of cells lacking ATM, suggesting a potential functional overlap between these proteins. ATRkd overexpression also causes increased sensitivity to hydroxyurea that is associated with microtubule-mediated nuclear abnormalities. These observations are consistent with uncoupling of certain mitotic events from the completion of S-phase. Thus, ATR is an important component of multiple DNA damage response pathways and may be involved in the DNA replication (S/M) checkpoint.

Full Text

The Full Text of this article is available as a PDF (292K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Allen JB, Zhou Z, Siede W, Friedberg EC, Elledge SJ. The SAD1/RAD53 protein kinase controls multiple checkpoints and DNA damage-induced transcription in yeast. Genes Dev. 1994 Oct 15;8(20):2401–2415. [PubMed]
  • Anderson HJ, Roberge M. DNA topoisomerase II: a review of its involvement in chromosome structure, DNA replication, transcription and mitosis. Cell Biol Int Rep. 1992 Aug;16(8):717–724. [PubMed]
  • Arlett CF, Green MH, Priestley A, Harcourt SA, Mayne LV. Comparative human cellular radiosensitivity: I. The effect of SV40 transformation and immortalisation on the gamma-irradiation survival of skin derived fibroblasts from normal individuals and from ataxia-telangiectasia patients and heterozygotes. Int J Radiat Biol. 1988 Dec;54(6):911–928. [PubMed]
  • Bentley NJ, Holtzman DA, Flaggs G, Keegan KS, DeMaggio A, Ford JC, Hoekstra M, Carr AM. The Schizosaccharomyces pombe rad3 checkpoint gene. EMBO J. 1996 Dec 2;15(23):6641–6651. [PMC free article] [PubMed]
  • Brown EJ, Schreiber SL. A signaling pathway to translational control. Cell. 1996 Aug 23;86(4):517–520. [PubMed]
  • Brown EJ, Beal PA, Keith CT, Chen J, Shin TB, Schreiber SL. Control of p70 s6 kinase by kinase activity of FRAP in vivo. Nature. 1995 Oct 5;377(6548):441–446. [PubMed]
  • Carr AM. Control of cell cycle arrest by the Mec1sc/Rad3sp DNA structure checkpoint pathway. Curr Opin Genet Dev. 1997 Feb;7(1):93–98. [PubMed]
  • Cimprich KA, Shin TB, Keith CT, Schreiber SL. cDNA cloning and gene mapping of a candidate human cell cycle checkpoint protein. Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):2850–2855. [PMC free article] [PubMed]
  • Clipstone NA, Crabtree GR. Identification of calcineurin as a key signalling enzyme in T-lymphocyte activation. Nature. 1992 Jun 25;357(6380):695–697. [PubMed]
  • Dasso M, Newport JW. Completion of DNA replication is monitored by a feedback system that controls the initiation of mitosis in vitro: studies in Xenopus. Cell. 1990 Jun 1;61(5):811–823. [PubMed]
  • Elledge SJ. Cell cycle checkpoints: preventing an identity crisis. Science. 1996 Dec 6;274(5293):1664–1672. [PubMed]
  • Enoch T, Nurse P. Mutation of fission yeast cell cycle control genes abolishes dependence of mitosis on DNA replication. Cell. 1990 Feb 23;60(4):665–673. [PubMed]
  • Enoch T, Carr AM, Nurse P. Fission yeast genes involved in coupling mitosis to completion of DNA replication. Genes Dev. 1992 Nov;6(11):2035–2046. [PubMed]
  • Gossen M, Bujard H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5547–5551. [PMC free article] [PubMed]
  • Gossen M, Freundlieb S, Bender G, Müller G, Hillen W, Bujard H. Transcriptional activation by tetracyclines in mammalian cells. Science. 1995 Jun 23;268(5218):1766–1769. [PubMed]
  • Greenwell PW, Kronmal SL, Porter SE, Gassenhuber J, Obermaier B, Petes TD. TEL1, a gene involved in controlling telomere length in S. cerevisiae, is homologous to the human ataxia telangiectasia gene. Cell. 1995 Sep 8;82(5):823–829. [PubMed]
  • Hall MN. The TOR signalling pathway and growth control in yeast. Biochem Soc Trans. 1996 Feb;24(1):234–239. [PubMed]
  • Hari KL, Santerre A, Sekelsky JJ, McKim KS, Boyd JB, Hawley RS. The mei-41 gene of D. melanogaster is a structural and functional homolog of the human ataxia telangiectasia gene. Cell. 1995 Sep 8;82(5):815–821. [PubMed]
  • Hartwell LH, Kastan MB. Cell cycle control and cancer. Science. 1994 Dec 16;266(5192):1821–1828. [PubMed]
  • Hawley RS, Friend SH. Strange bedfellows in even stranger places: the role of ATM in meiotic cells, lymphocytes, tumors, and its functional links to p53. Genes Dev. 1996 Oct 1;10(19):2383–2388. [PubMed]
  • Hoekstra MF. Responses to DNA damage and regulation of cell cycle checkpoints by the ATM protein kinase family. Curr Opin Genet Dev. 1997 Apr;7(2):170–175. [PubMed]
  • Jackson SP. DNA damage detection by DNA dependent protein kinase and related enzymes. Cancer Surv. 1996;28:261–279. [PubMed]
  • Jackson SP. The recognition of DNA damage. Curr Opin Genet Dev. 1996 Feb;6(1):19–25. [PubMed]
  • Jimenez G, Yucel J, Rowley R, Subramani S. The rad3+ gene of Schizosaccharomyces pombe is involved in multiple checkpoint functions and in DNA repair. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4952–4956. [PMC free article] [PubMed]
  • Kastan MB. Signalling to p53: where does it all start? Bioessays. 1996 Aug;18(8):617–619. [PubMed]
  • Kastan MB, Zhan Q, el-Deiry WS, Carrier F, Jacks T, Walsh WV, Plunkett BS, Vogelstein B, Fornace AJ., Jr A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia. Cell. 1992 Nov 13;71(4):587–597. [PubMed]
  • Kato R, Ogawa H. An essential gene, ESR1, is required for mitotic cell growth, DNA repair and meiotic recombination in Saccharomyces cerevisiae. Nucleic Acids Res. 1994 Aug 11;22(15):3104–3112. [PMC free article] [PubMed]
  • Keegan KS, Holtzman DA, Plug AW, Christenson ER, Brainerd EE, Flaggs G, Bentley NJ, Taylor EM, Meyn MS, Moss SB, et al. The Atr and Atm protein kinases associate with different sites along meiotically pairing chromosomes. Genes Dev. 1996 Oct 1;10(19):2423–2437. [PubMed]
  • Keith CT, Schreiber SL. PIK-related kinases: DNA repair, recombination, and cell cycle checkpoints. Science. 1995 Oct 6;270(5233):50–51. [PubMed]
  • Kinzler KW, Vogelstein B. Lessons from hereditary colorectal cancer. Cell. 1996 Oct 18;87(2):159–170. [PubMed]
  • Kumagai A, Dunphy WG. Control of the Cdc2/cyclin B complex in Xenopus egg extracts arrested at a G2/M checkpoint with DNA synthesis inhibitors. Mol Biol Cell. 1995 Feb;6(2):199–213. [PMC free article] [PubMed]
  • Levine AJ. p53, the cellular gatekeeper for growth and division. Cell. 1997 Feb 7;88(3):323–331. [PubMed]
  • Lew DJ, Kornbluth S. Regulatory roles of cyclin dependent kinase phosphorylation in cell cycle control. Curr Opin Cell Biol. 1996 Dec;8(6):795–804. [PubMed]
  • Linke SP, Clarkin KC, Di Leonardo A, Tsou A, Wahl GM. A reversible, p53-dependent G0/G1 cell cycle arrest induced by ribonucleotide depletion in the absence of detectable DNA damage. Genes Dev. 1996 Apr 15;10(8):934–947. [PubMed]
  • Lydall D, Weinert T. From DNA damage to cell cycle arrest and suicide: a budding yeast perspective. Curr Opin Genet Dev. 1996 Feb;6(1):4–11. [PubMed]
  • Meyn MS. Ataxia-telangiectasia and cellular responses to DNA damage. Cancer Res. 1995 Dec 15;55(24):5991–6001. [PubMed]
  • Morrow DM, Tagle DA, Shiloh Y, Collins FS, Hieter P. TEL1, an S. cerevisiae homolog of the human gene mutated in ataxia telangiectasia, is functionally related to the yeast checkpoint gene MEC1. Cell. 1995 Sep 8;82(5):831–840. [PubMed]
  • Nevins JR. Cell cycle targets of the DNA tumor viruses. Curr Opin Genet Dev. 1994 Feb;4(1):130–134. [PubMed]
  • Painter RB, Young BR. Radiosensitivity in ataxia-telangiectasia: a new explanation. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7315–7317. [PMC free article] [PubMed]
  • Paules RS, Levedakou EN, Wilson SJ, Innes CL, Rhodes N, Tlsty TD, Galloway DA, Donehower LA, Tainsky MA, Kaufmann WK. Defective G2 checkpoint function in cells from individuals with familial cancer syndromes. Cancer Res. 1995 Apr 15;55(8):1763–1773. [PubMed]
  • Paulovich AG, Hartwell LH. A checkpoint regulates the rate of progression through S phase in S. cerevisiae in response to DNA damage. Cell. 1995 Sep 8;82(5):841–847. [PubMed]
  • Paulovich AG, Toczyski DP, Hartwell LH. When checkpoints fail. Cell. 1997 Feb 7;88(3):315–321. [PubMed]
  • Perlmutter RM, Alberola-Ila J. The use of dominant-negative mutations to elucidate signal transduction pathways in lymphocytes. Curr Opin Immunol. 1996 Apr;8(2):285–290. [PubMed]
  • Prickett KS, Amberg DC, Hopp TP. A calcium-dependent antibody for identification and purification of recombinant proteins. Biotechniques. 1989 Jun;7(6):580–589. [PubMed]
  • Rudner AD, Murray AW. The spindle assembly checkpoint. Curr Opin Cell Biol. 1996 Dec;8(6):773–780. [PubMed]
  • Sanchez Y, Desany BA, Jones WJ, Liu Q, Wang B, Elledge SJ. Regulation of RAD53 by the ATM-like kinases MEC1 and TEL1 in yeast cell cycle checkpoint pathways. Science. 1996 Jan 19;271(5247):357–360. [PubMed]
  • Savitsky K, Sfez S, Tagle DA, Ziv Y, Sartiel A, Collins FS, Shiloh Y, Rotman G. The complete sequence of the coding region of the ATM gene reveals similarity to cell cycle regulators in different species. Hum Mol Genet. 1995 Nov;4(11):2025–2032. [PubMed]
  • Schmidt A, Kunz J, Hall MN. TOR2 is required for organization of the actin cytoskeleton in yeast. Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13780–13785. [PMC free article] [PubMed]
  • Seaton BL, Yucel J, Sunnerhagen P, Subramani S. Isolation and characterization of the Schizosaccharomyces pombe rad3 gene, involved in the DNA damage and DNA synthesis checkpoints. Gene. 1992 Sep 21;119(1):83–89. [PubMed]
  • Siede W, Allen JB, Elledge SJ, Friedberg EC. The Saccharomyces cerevisiae MEC1 gene, which encodes a homolog of the human ATM gene product, is required for G1 arrest following radiation treatment. J Bacteriol. 1996 Oct;178(19):5841–5843. [PMC free article] [PubMed]
  • Stewart E, Enoch T. S-phase and DNA-damage checkpoints: a tale of two yeasts. Curr Opin Cell Biol. 1996 Dec;8(6):781–787. [PubMed]
  • Stewart E, Chapman CR, Al-Khodairy F, Carr AM, Enoch T. rqh1+, a fission yeast gene related to the Bloom's and Werner's syndrome genes, is required for reversible S phase arrest. EMBO J. 1997 May 15;16(10):2682–2692. [PMC free article] [PubMed]
  • Weaver DT. What to do at an end: DNA double-strand-break repair. Trends Genet. 1995 Oct;11(10):388–392. [PubMed]
  • Weinert TA, Kiser GL, Hartwell LH. Mitotic checkpoint genes in budding yeast and the dependence of mitosis on DNA replication and repair. Genes Dev. 1994 Mar 15;8(6):652–665. [PubMed]
  • Zheng XF, Florentino D, Chen J, Crabtree GR, Schreiber SL. TOR kinase domains are required for two distinct functions, only one of which is inhibited by rapamycin. Cell. 1995 Jul 14;82(1):121–130. [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...