Skip to main content
NCBI home page
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 2001 Jun;92(6):710–718. doi: 10.1111/j.1349-7006.2001.tb01152.x

X‐Irradiation Induces Up‐regulation of ATM Gene Expression in Wild‐type Lymphoblastoid Cell Lines, but Not in Their Heterozygous or Homozygous Ataxia‐telangiectasia Counterparts

Yuko Hirai 1,✉, Tomonori Hayashi 1, Yoshiko Kubo 1, Yuko Hoki 2, Izumi Arita 3, Kouichi Tatsumi 2, Toshio Seyama 1
PMCID: PMC5926759  PMID: 11429062

Abstract

Ataxia‐telangiectasia (AT) is an autosomal recessive disease. The relevant gene has been cloned and designated ATM. We studied the expression of both ATM mRNA and the ATM protein in unirradi‐ated and X‐irradiated EBV (Epstein‐Barr virus)‐transformed lymphoblastoid cell lines (LCLs) derived from donors who were normal (ATM+/+), AT heterozygotes (ATM+/−), or AT homozy‐gotes (ATM−/−), respectively. In ATM+/+ LCLs, the levels of ATM mRNA were found to have increased by approximately 1.5‐fold within 1 h of exposure to 10 Gy of X‐rays, while the ATM protein levels had increased by 1.5‐ to 2.0‐fold within 2 to 3 h of irradiation. The wild‐type mRNA and protein levels both returned to their basal values fairly quickly after this tune. The results obtained with the ATM+/− LCLs were quite different, however: neither the mRNA nor protein levels were found to have increased as a consequence of X‐irradiation in any ATM+/− LCL. Twelve of the mutations in the ATM−/− LCLs we used were truncating mutations, and we suspected that the corresponding truncated ATM proteins would be too labile to be detected by western blot analysis. However, five of the ATM−/− LCLs produced mutant ATM proteins that were identical in molecular weight to the wild‐type ATM protein. When cells from three of these five clones were exposed to X‐rays, transcription of the mutant ATM genes appeared to reduce somewhat, as were the levels of protein being produced. These results suggest that the normal ATM gene responds to ionizing radiation by up‐regulating its activity, whereas none of the mutant ATM genes we studied were able to respond in this way.

Keywords: Ataxia, telangiectasia (AT), Irradiation, ATM expression, Up, regulation, Lymphoblastoid cell lines (LCLs)

Full Text

The Full Text of this article is available as a PDF (245.1 KB).

Reference

  • 1.Harnden , D. G.The nature of ataxia‐telangiectasia: problems and perspectives . Int. J. Radial. Biol , 66 , S13 – S19 ( 1994. ). [PubMed] [Google Scholar]
  • 2.Gatti , R. A. , Boder , E. , Vinters , H. V. , Sparkes , R. S. , Norman , A. and Lange , K.Ataxia‐telangiectasia: an interdisciplinary approach to pathogenesis . Medicine , 70 , 99 – 117 ( 1991. ). [PubMed] [Google Scholar]
  • 3.Taylor , A. M. , Harnden , D. G. , Arlett , C. F. , Harcourt , S. A. , Lehmann , A. R. , Stevens , S. and Bridges , B. A.Ataxia telangiectasia: a human mutation with abnormal radiation sensitivity . Nature , 258 , 427 – 429 ( 1975. ). [DOI] [PubMed] [Google Scholar]
  • 4.Young , B. R. and Painter , R. B.Radioresistant DNA synthesis and human genetic diseases . Hum. Genet. , 82 , 113 – 117 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 5.Beamish , H. and Lavin , M. F.Radiosensitivity in ataxia‐telangiectasia: anomalies in radiation‐induced cell cycle delay . Int. J. Radial. Biol , 65 , 175 – 184 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 6.Savitsky , K. , Bar‐Shira , A. , Gilad , S. , Rotman , G. , Ziv , Y. , Vanagaite , L. , Tagle , D. A. , Smith , S. , Uziel , T. , Sfez , S. and Shiloh , Y.A single ataxia telangiectasia gene with a product similar to PI‐3 kinase . Science , 268 , 1749 – 1753 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 7.Savitsky , K. , Sfez , S. , Tagle , D. A. , Ziv , Y. , Sartiel , A. , Collins , F. S. , Shiloh , Y. and 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. , 4 , 2025 – 2032 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 8.Gilad , S. , Khosravi , R. , Shkedy , D. , Uziel , T. , Ziv , Y. , Savitsky , K. , Rotman , G. , Smith , S. , Chessa , L. , Jorgensen , T. J. , Harnik , R. , Frydman , M. , Sanal , O. , Portnoi , S. , Goldwicz , Z. , Jaspers , N. G. , Gatti , R. A. , Lenoir , G. , Lavin , M. F. , Tatsumi , K. , Wegner , R. D. , Shiloh , Y. and Bar‐Shira , A.Predominance of null mutations in ataxia‐telangiectasia . Hum. Mol. Genet. , 5 , 433 – 439 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 9.Meyn , M. S.Ataxia‐telangiectasia and cellular responses to DNA damage . Cancer Res. , 55 , 5991 – 6001 ( 1995. ). [PubMed] [Google Scholar]
  • 10.Beamish , H. , Williams , R. , Chen , P. and Lavin , M. F.Defect in multiple cell cycle checkpoints in ataxia‐telangiectasia postirradiation . J. BioL Chem. , 271 , 20486 – 20493 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 11.Lavin , M. F. and Shiloh , Y.The genetic defect in ataxia‐telangiectasia . Annu. Rev. Immunol. , 15 , 177 – 202 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 12.Khanna , K. K. , Keating , K. E. , Kozlov , S. , Scott , S. , Gatei , M. , Hobson , K. , Taya , Y. , Gabrielli , B. , Chan , D. , Lees‐Miller , S. P. and Lavin , M. F.ATM associates with and phosphorylates p53: mapping the region of interaction . Nat. Genet. , 20 , 398 – 400 ( 1998. ). [DOI] [PubMed] [Google Scholar]
  • 13.Siliciano , J. D. , Canman , C. E. , Taya , Y. , Sakaguchi , K. , Appella , E. and Kastan , M. B.DNA damage induces phosphorylation of the amino terminus of p53 . Genes Dev. , 11 , 3471 – 3481 ( 1997. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Banin , S. , Moyal , L. , Shieh , S. , Taya , Y. , Anderson , C. W. , Chessa , L. , Smorodinsky , N. L , Prives , C. , Reiss , Y. , Shiloh , Y. and Ziv , Y.Enhanced phosphorylation of p53 by ATM in response to DNA damage . Science , 281 , 1674 – 1677 ( 1998. ). [DOI] [PubMed] [Google Scholar]
  • 15.Canman , C. E. , Lim , D. S. , Cimprich , K. A. , Taya , Y. , Tamai , K. , Sakaguchi , K. , Appella , E. , Kastan , M. B. and Siliciano , J. D.Activation of the ATM kinase by ionizing radiation and phosphorylation of p53 . Science , 281 , 1677 – 1679 ( 1998. ). [DOI] [PubMed] [Google Scholar]
  • 16.Walters , D. , Khanna , K. K. , Beamish , H. , Birrell , G. , Spring , K. , Kedar , P. , Gatei , M. , Stenzel , D. , Hobson , K. , Kozlov , S. , Zhang , N. , Farrell , A. , Ramsay , J. , Gatti , R. and Lavin , M.Cellular localisation of the ataxia‐telangiectasia (ATM) gene product and discrimination between mutated and normal forms . Oncogens , 14 , 1911 – 1921 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 17.Lakin , N. D. , Weber , P. , Stankovic , T. , Rottinghaus , S. T. , Taylor , A. M. and Jackson , S. P . Analysis of the ATM protein in wild‐type and ataxia telangiectasia cells . Oncogene , 13 , 2707 – 2716 ( 1996. ). [PubMed] [Google Scholar]
  • 18.Brown , K. D. , Ziv , Y. , Sadanandan , S. N. , Chessa , L. , Collins , F. S. , Shiloh , Y. and Tagle , D.A. The ataxia‐telangiectasia gene product, a constitutively expressed nuclear protein that is not up‐regulated following genome damage . Proc. Natl. Acad. Sci. USA , 94 , 1840 – 1845 ( 1997. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Pandita , T. K. , Lieberman , H. B. , Lim , D. S. , Dhar , S. , Zheng , W. , Taya , Y. and Kastan , M. B.Ionizing radiation activates the ATM kinase throughout the cell cycle . Oncogene , 19 , 1386 – 1391 ( 2000. ). [DOI] [PubMed] [Google Scholar]
  • 20.Arita , I. , Tachibana , A. , Takebe , H. and Tatsumi , K.Predominance of Mex+ cells in newly‐established human lym‐phoblastoid cell lines . Carcinogenesis , 10 , 2067 – 2073 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 21.Wright , J. , Teraoka , S. , Onengut , S. , Tolun , A. , Gatti , R. A. , Ochs , H. D.Wright , J. , Teraoka , S. , Onengut , S. , Tolun , A. , Gatti , R. A. , Ochs , H. D. and Concannon , P. Ahigh frequency of distinct ATM gene mutations in ataxia‐telangiectasia . Am. J. Hum. Genet. , 59 , 839 – 846 ( 1996. ). [PMC free article] [PubMed] [Google Scholar]
  • 22.Ejima , Y. and Sasaki , M. S.Mutations of the ATM gene detected in Japanese ataxia‐telangiectasia patients: possible preponderance of the two founder mutations 4612dell65 and 7883del5 . Hum. Genet , 102 , 403 – 408 ( 1998. ). [DOI] [PubMed] [Google Scholar]
  • 23.Harlow , E. and Lane , D. In “ Antibodies. A Laboratory Manual ” ( 1988. ). Cold Spring Harbor Laboratory; , New York . [Google Scholar]
  • 24.Inoue , A. , Miki , S. , Seto , M. , Kikuchi , T. , Morita , S. , Ueda , H. , Misu , Y. and Nakata , Y.Aripiprazole, a novel antipsy‐chotic drug, inhibits quinpirole‐evoked GTPase activity but does not up‐regulate dopamine D2 receptor following repeated treatment in the rat striatum . Eur. J. Pharmacol. , 321 , 105 – 111 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 25.Uziel , T. , Savitsky , K. , Platzer , M. , Ziv , Y. , Helbitz , T. , Nehls , M. , Boehm , T. , Rosenthal , A. , Shiloh , Y. and Rotman , G.Genomic organization of the ATM gene . Genomics , 33 , 317 – 320 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 26.Krause , J. E. , Cremins , J. D. , Carter , M. S. , Brown , E. R. and MacDonald , M. R . Solution hybridization‐nuclease protection assays for sensitive detection of differentially spliced substance P‐ and neurokinin A‐encoding messenger ribonucleic acids . Methods Enzymol. , 168 , 634 – 652 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 27.Fukao , T. , Kaneko , H. , Birrell , G. , Gatei , M. , Tashita , H. , Yoshida , T. , Cross , S. , Kedar , P. , Walters , D. , Khana , K. K. , Misko , I. , Kondo , N. and Lavin , M. F.ATM is upregulated during The milogenic response in peripheral blood mononuclear cells . Blood , 94 , 1998 – 2006 ( 1999. ). [PubMed] [Google Scholar]
  • 28.Byrd , P. J. , Cooper , P. R. , Stankovic , T. , Kullar , H. S. , Walls , G. D. , Robinson , P. J. and Taylor , M. R.A gene transcribed from the bidirectional ATM promoter coding for a serine rich protein: amino acid sequence, structure and expression studies . Hum. Mol. Genet. , 5 , 1785 – 1791 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 29.Imai , T. , Yamauchi , M. , Seki , N. , Sugawara , T. , Saito , T. , Matsuda , Y. , Ito , H. , Nagase , T. , Nomura , N. and Hori , T.Identification and characterization of a new gene physically linked to the ATM gene . Genome Res. , 6 , 439 – 447 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 30.Imai , T. , Sugawara , T. , Nishiyama , A. , Shimada , R. , Ohki , R. , Seki , N. , Sagara , M. , Ito , H. , Yamauchi , M. and Hori , T.The structure and organization of the human NPAT gene . Genomics , 42 , 388 – 392 ( 1997. ). [DOI] [PubMed] [Google Scholar]
  • 31.Gueven , N. , Keating , K. E. , Chen , P. , Fukao , T. , Khanna , K. K. , Wallers , D. , Rodemann , P. H. and Lavin , M. F . EOF sensitizes cells lo ionizing radiation by downregulating ATM . J. Biol. Chem. , 276 , 8884 – 8891 ( 2001. ). [DOI] [PubMed] [Google Scholar]

Articles from Japanese Journal of Cancer Research : Gann are provided here courtesy of Wiley

RESOURCES