Two essential but distinct functions of the mammalian abasic endonuclease

April 11, 2005
102 (16) 5739-5743

Abstract

The mammalian abasic endonuclease, APE1, has two distinct roles in the repair of oxidative DNA damage and in gene regulation. Here we show that both functions are essential for cell survival. Deletion of the APE1 gene causes embryonic lethality in mice, and no nullizygous embryo fibroblasts have been isolated. We have now established nullizygous embryo fibroblast lines from APE1–/– mouse embryos that are transgenic with the “floxed” human APE1 (hAPE1) gene. Removal of hAPE1 by Cre expression through nuclear microinjection elicited apoptosis in these cells within 24 h, which was blocked by coinjection of the wild-type hAPE1 gene. In contrast, mutant hAPE1 alleles, lacking either the DNA repair or acetylation-mediated gene regulatory function, could not prevent apoptosis, although the combination of these two mutants complemented APE deficiency induced by Cre. These results indicate that distinct and separable functions of APE1 are both essential for mammalian cells even in vitro and provide the evidence that mammalian cells, unlike yeast or Escherichia coli, absolutely require APE for survival, presumably to protect against spontaneous oxidative DNA damage.

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Acknowledgments

We thank Mr. E. F. Willmore, Jr., for animal husbandry and excellent technical assistance; Dr. B. Sauer for the Cre-EGFP fusion vector, Dr. C. Yallampalli for guidance in embryo isolation; Dr. J. Ceci for generating the APE1 transgenic mice; Drs. D. Konkel, M. Weinfeld, and S. Adhya for critical reading of the manuscript; and Ms. W. Smith for secretarial help. This research was supported by U.S. Public Health Service Grants R01 ES08457, R01 CA53791, P01 06676, and R01 CA98664.

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Information & Authors

Information

Published in

Go to Proceedings of the National Academy of Sciences
Go to Proceedings of the National Academy of Sciences
Proceedings of the National Academy of Sciences
Vol. 102 | No. 16
April 19, 2005
PubMed: 15824325

Classifications

Submission history

Received: December 9, 2004
Published online: April 11, 2005
Published in issue: April 19, 2005

Keywords

  1. conditional gene inactivation
  2. DNA repair
  3. endogenous DNA damage
  4. base excision repair

Acknowledgments

We thank Mr. E. F. Willmore, Jr., for animal husbandry and excellent technical assistance; Dr. B. Sauer for the Cre-EGFP fusion vector, Dr. C. Yallampalli for guidance in embryo isolation; Dr. J. Ceci for generating the APE1 transgenic mice; Drs. D. Konkel, M. Weinfeld, and S. Adhya for critical reading of the manuscript; and Ms. W. Smith for secretarial help. This research was supported by U.S. Public Health Service Grants R01 ES08457, R01 CA53791, P01 06676, and R01 CA98664.

Authors

Affiliations

Tadahide Izumi
Sealy Center for Molecular Science, Department of Human Biological Chemistry and Genetics, and Department of Surgery, University of Texas Medical Branch, Galveston, TX 77555; Biosciences Division, Los Alamos National Laboratory, Los Alamos, NM 87545; and Department of Radiation Oncology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390
David B. Brown
Sealy Center for Molecular Science, Department of Human Biological Chemistry and Genetics, and Department of Surgery, University of Texas Medical Branch, Galveston, TX 77555; Biosciences Division, Los Alamos National Laboratory, Los Alamos, NM 87545; and Department of Radiation Oncology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390
C. V. Naidu§
Sealy Center for Molecular Science, Department of Human Biological Chemistry and Genetics, and Department of Surgery, University of Texas Medical Branch, Galveston, TX 77555; Biosciences Division, Los Alamos National Laboratory, Los Alamos, NM 87545; and Department of Radiation Oncology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390
Kishor K. Bhakat
Sealy Center for Molecular Science, Department of Human Biological Chemistry and Genetics, and Department of Surgery, University of Texas Medical Branch, Galveston, TX 77555; Biosciences Division, Los Alamos National Laboratory, Los Alamos, NM 87545; and Department of Radiation Oncology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390
Mark A. MacInnes
Sealy Center for Molecular Science, Department of Human Biological Chemistry and Genetics, and Department of Surgery, University of Texas Medical Branch, Galveston, TX 77555; Biosciences Division, Los Alamos National Laboratory, Los Alamos, NM 87545; and Department of Radiation Oncology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390
Hiroshi Saito
Sealy Center for Molecular Science, Department of Human Biological Chemistry and Genetics, and Department of Surgery, University of Texas Medical Branch, Galveston, TX 77555; Biosciences Division, Los Alamos National Laboratory, Los Alamos, NM 87545; and Department of Radiation Oncology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390
David J. Chen
Sealy Center for Molecular Science, Department of Human Biological Chemistry and Genetics, and Department of Surgery, University of Texas Medical Branch, Galveston, TX 77555; Biosciences Division, Los Alamos National Laboratory, Los Alamos, NM 87545; and Department of Radiation Oncology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390
Sankar Mitra††
Sealy Center for Molecular Science, Department of Human Biological Chemistry and Genetics, and Department of Surgery, University of Texas Medical Branch, Galveston, TX 77555; Biosciences Division, Los Alamos National Laboratory, Los Alamos, NM 87545; and Department of Radiation Oncology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390

Notes

††
To whom correspondence should be addressed. E-mail: [email protected].
Present address: Health Sciences Center, Louisiana State University, New Orleans, LA 70112.
§
Present address: Department of Biotechnology, Sri Venkateswara University, Tirupati 517502, India.
Communicated by Richard B. Setlow, Brookhaven National Laboratory, Upton, NY, February 16, 2005

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    Two essential but distinct functions of the mammalian abasic endonuclease
    Proceedings of the National Academy of Sciences
    • Vol. 102
    • No. 16
    • pp. 5637-5897

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