Congenital Bone Marrow Failure in DNA-PKcs Mutant Mice Associated with Deficiencies in DNA Repair

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 2011 Apr 13

PMID 21482716

Citations 87

Authors

Shichuan Zhang

Hirohiko Yajima

HoangDinh Huynh

Junke Zheng

Elsa Callen

Hua-Tang Chen

Nancy Wong

Samuel Bunting

Yu-Fen Lin

Mengxia Li

Kyung-Jone Lee

Michael Story

Eric Gapud

Barry P Sleckman

Andre Nussenzweig

Cheng Cheng Zhang

David J Chen

Benjamin P C Chen

Affiliations

Soon will be listed here.

Abstract

The nonhomologous end-joining (NHEJ) pathway is essential for radioresistance and lymphocyte-specific V(D)J (variable [diversity] joining) recombination. Defects in NHEJ also impair hematopoietic stem cell (HSC) activity with age but do not affect the initial establishment of HSC reserves. In this paper, we report that, in contrast to deoxyribonucleic acid (DNA)-dependent protein kinase catalytic subunit (DNA-PKcs)-null mice, knockin mice with the DNA-PKcs(3A/3A) allele, which codes for three alanine substitutions at the mouse Thr2605 phosphorylation cluster, die prematurely because of congenital bone marrow failure. Impaired proliferation of DNA-PKcs(3A/3A) HSCs is caused by excessive DNA damage and p53-dependent apoptosis. In addition, increased apoptosis in the intestinal crypt and epidermal hyperpigmentation indicate the presence of elevated genotoxic stress and p53 activation. Analysis of embryonic fibroblasts further reveals that DNA-PKcs(3A/3A) cells are hypersensitive to DNA cross-linking agents and are defective in both homologous recombination and the Fanconi anemia DNA damage response pathways. We conclude that phosphorylation of DNA-PKcs is essential for the normal activation of multiple DNA repair pathways, which in turn is critical for the maintenance of diverse populations of tissue stem cells in mice.

Citing Articles

DNA-PK controls Apollo's access to leading-end telomeres.

Sonmez C, Toia B, Eickhoff P, Matei A, El Beyrouthy M, Wallner B Nucleic Acids Res. 2024; 52(8):4313-4327.

PMID: 38407308 PMC: 11077071. DOI: 10.1093/nar/gkae105.

Targeting DNA-PK.

Novotny J, Marino-Enriquez A, Fletcher J Cancer Treat Res. 2023; 186:299-312.

PMID: 37978142 PMC: 11870302. DOI: 10.1007/978-3-031-30065-3_16.

LepR+ niche cell-derived AREG compromises hematopoietic stem cell maintenance under conditions of DNA repair deficiency and aging.

Wu L, Lin Q, Chatla S, Amarachintha S, Wilson A, Atale N Blood. 2023; 142(18):1529-1542.

PMID: 37584437 PMC: 10656728. DOI: 10.1182/blood.2022018212.

Genotoxic aldehyde stress prematurely ages hematopoietic stem cells in a p53-driven manner.

Wang M, Brandt L, Wang X, Russell H, Mitchell E, Kamimae-Lanning A Mol Cell. 2023; 83(14):2417-2433.e7.

PMID: 37348497 PMC: 7614878. DOI: 10.1016/j.molcel.2023.05.035.

Phosphorylation of DNA-PKcs at the S2056 cluster ensures efficient and productive lymphocyte development in XLF-deficient mice.

Zhu Y, Jiang W, Lee B, Li A, Gershik S, Zha S Proc Natl Acad Sci U S A. 2023; 120(25):e2221894120.

PMID: 37307443 PMC: 10288554. DOI: 10.1073/pnas.2221894120.

References

Prasher J, Lalai A, Heijmans-Antonissen C, Ploemacher R, Hoeijmakers J, Touw I . Reduced hematopoietic reserves in DNA interstrand crosslink repair-deficient Ercc1-/- mice. EMBO J. 2005; 24(4):861-71. PMC: 549615. DOI: 10.1038/sj.emboj.7600542. View

Kondo M, Wagers A, Manz M, Prohaska S, Scherer D, Beilhack G . Biology of hematopoietic stem cells and progenitors: implications for clinical application. Annu Rev Immunol. 2003; 21:759-806. DOI: 10.1146/annurev.immunol.21.120601.141007. View

Difilippantonio S, Gapud E, Wong N, Huang C, Mahowald G, Chen H . 53BP1 facilitates long-range DNA end-joining during V(D)J recombination. Nature. 2008; 456(7221):529-33. PMC: 3596817. DOI: 10.1038/nature07476. View

Hockemeyer D, Palm W, Wang R, Couto S, de Lange T . Engineered telomere degradation models dyskeratosis congenita. Genes Dev. 2008; 22(13):1773-85. PMC: 2492664. DOI: 10.1101/gad.1679208. View

Bunting S, Callen E, Wong N, Chen H, Polato F, Gunn A . 53BP1 inhibits homologous recombination in Brca1-deficient cells by blocking resection of DNA breaks. Cell. 2010; 141(2):243-54. PMC: 2857570. DOI: 10.1016/j.cell.2010.03.012. View

Moldovan G, DAndrea A . How the fanconi anemia pathway guards the genome. Annu Rev Genet. 2009; 43:223-49. PMC: 2830711. DOI: 10.1146/annurev-genet-102108-134222. View

Taccioli G, Amatucci A, Beamish H, Gell D, Xiang X, Torres Arzayus M . Targeted disruption of the catalytic subunit of the DNA-PK gene in mice confers severe combined immunodeficiency and radiosensitivity. Immunity. 1998; 9(3):355-66. DOI: 10.1016/s1074-7613(00)80618-4. View

Wang Y, Schulte B, LaRue A, Ogawa M, Zhou D . Total body irradiation selectively induces murine hematopoietic stem cell senescence. Blood. 2005; 107(1):358-66. PMC: 1895367. DOI: 10.1182/blood-2005-04-1418. View

Bender C, Sikes M, Sullivan R, Huye L, Le Beau M, Roth D . Cancer predisposition and hematopoietic failure in Rad50(S/S) mice. Genes Dev. 2002; 16(17):2237-51. PMC: 186667. DOI: 10.1101/gad.1007902. View

10.

Ding Q, Reddy Y, Wang W, Woods T, Douglas P, Ramsden D . Autophosphorylation of the catalytic subunit of the DNA-dependent protein kinase is required for efficient end processing during DNA double-strand break repair. Mol Cell Biol. 2003; 23(16):5836-48. PMC: 166339. DOI: 10.1128/MCB.23.16.5836-5848.2003. View

11.

Mirchandani K, DAndrea A . The Fanconi anemia/BRCA pathway: a coordinator of cross-link repair. Exp Cell Res. 2006; 312(14):2647-53. DOI: 10.1016/j.yexcr.2006.06.014. View

12.

Weterings E, Chen D . The endless tale of non-homologous end-joining. Cell Res. 2008; 18(1):114-24. DOI: 10.1038/cr.2008.3. View

13.

Sonoda E, Hochegger H, Saberi A, Taniguchi Y, Takeda S . Differential usage of non-homologous end-joining and homologous recombination in double strand break repair. DNA Repair (Amst). 2006; 5(9-10):1021-9. DOI: 10.1016/j.dnarep.2006.05.022. View

14.

Kurimasa A, Ouyang H, Dong L, Wang S, Li X, Cordon-Cardo C . Catalytic subunit of DNA-dependent protein kinase: impact on lymphocyte development and tumorigenesis. Proc Natl Acad Sci U S A. 1999; 96(4):1403-8. PMC: 15475. DOI: 10.1073/pnas.96.4.1403. View

15.

Meek K, Douglas P, Cui X, Ding Q, Lees-Miller S . trans Autophosphorylation at DNA-dependent protein kinase's two major autophosphorylation site clusters facilitates end processing but not end joining. Mol Cell Biol. 2007; 27(10):3881-90. PMC: 1899996. DOI: 10.1128/MCB.02366-06. View

16.

Adamo A, Collis S, Adelman C, Silva N, Horejsi Z, Ward J . Preventing nonhomologous end joining suppresses DNA repair defects of Fanconi anemia. Mol Cell. 2010; 39(1):25-35. DOI: 10.1016/j.molcel.2010.06.026. View

17.

Nacht M, Strasser A, Chan Y, Harris A, Schlissel M, Bronson R . Mutations in the p53 and SCID genes cooperate in tumorigenesis. Genes Dev. 1996; 10(16):2055-66. DOI: 10.1101/gad.10.16.2055. View

18.

Garcia-Higuera I, Taniguchi T, Ganesan S, Meyn M, Timmers C, Hejna J . Interaction of the Fanconi anemia proteins and BRCA1 in a common pathway. Mol Cell. 2001; 7(2):249-62. DOI: 10.1016/s1097-2765(01)00173-3. View

19.

McGowan K, Li J, Park C, Beaudry V, Tabor H, Sabnis A . Ribosomal mutations cause p53-mediated dark skin and pleiotropic effects. Nat Genet. 2008; 40(8):963-70. PMC: 3979291. DOI: 10.1038/ng.188. View

20.

Manis J, Dudley D, Kaylor L, Alt F . IgH class switch recombination to IgG1 in DNA-PKcs-deficient B cells. Immunity. 2002; 16(4):607-17. DOI: 10.1016/s1074-7613(02)00306-0. View