Skip to main content
OSTI.GOVU.S. Department of Energy Office of Scientific and Technical Information
U.S. Department of Energy
Office of Scientific and Technical Information
 

Restoration of G1 chemo/radioresistance and double-strand-break repair proficiency by wild-type but not endonuclease-deficient Artemis

Journal Article · · Nucleic Acids Research
DOI:https://doi.org/10.1093/nar/gkr257· OSTI ID:1625488
 [1];  [2];  [2];  [2];  [3]
  1. Virginia Commonwealth Univ., Richmond, VA (United States); DOE/OSTI
  2. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  3. Virginia Commonwealth Univ., Richmond, VA (United States)
+ Show Author Affiliations

Deficiency in Artemis is associated with lack of V(D)J recombination, sensitivity to radiation and radiomimetic drugs, and failure to repair a subset of DNA double-strand breaks (DSBs). Artemis harbors an endonuclease activity that trims both 5' - and 3' -ends of DSBs. To examine whether endonucleolytic trimming of terminally blocked DSBs by Artemis is a biologically relevant function, Artemis deficient fibroblasts were stably complemented with either wild-type Artemis or an endonuclease deficient D165N mutant. Wild-type Artemis completely restored resistance to $$\gamma$$-rays, bleomycin and neocarzinostatin, and also restored DSB-repair proficiency in G0/G1 phase as measured by pulsed field gel electrophoresis and repair focus resolution. In contrast, cells expressing the D165N mutant, even at very high levels, remained as chemo/ radiosensitive and repair deficient as the parental cells, as evidenced by persistent $$\gamma$$-H2AX, 53BP1 and Mre11 foci that slowly increased in size and ultimately became juxtaposed with promyelocytic leukemia protein nuclear bodies. In normal fibroblasts, overexpression of wild-type Artemis increased radio resistance, while D165N overexpression conferred partial repair deficiency following high dose radiation. Restoration of chemo/radio resistance by wild-type, but not D165N Artemis suggests that the lack of endonucleolytic trimming of DNA ends is the principal cause of sensitivity to double strand cleaving agents in Artemis-deficient cells.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division; National Cancer Institute (NCI); National Institutes of Health (NIH)
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
1625488
Journal Information:
Nucleic Acids Research, Journal Name: Nucleic Acids Research Journal Issue: 15 Vol. 39; ISSN 0305-1048
Publisher:
Oxford University PressCopyright Statement
Country of Publication:
United States
Language:
English

References (48)

DNA damage and mutagenesis by radiomimetic DNA-cleaving agents: bleomycin, neocarzinostatin and other enediynes journal August 1996
Retinoic acid regulates aberrant nuclear localization of PML-RARα in acute promyelocytic leukemia cells journal January 1994
DNA-PKcs regulates a single-stranded DNA endonuclease activity of Artemis journal April 2010
Purification and characterization of exonuclease-free Artemis: Implications for DNA-PK-dependent processing of DNA termini in NHEJ-catalyzed DSB repair journal June 2010
ATM Breaks into Heterochromatin journal August 2008
Artemis, a Novel DNA Double-Strand Break Repair/V(D)J Recombination Protein, Is Mutated in Human Severe Combined Immune Deficiency journal April 2001
Hairpin Opening and Overhang Processing by an Artemis/DNA-Dependent Protein Kinase Complex in Nonhomologous End Joining and V(D)J Recombination journal March 2002
GC/MS Methods To Quantify the 2-Deoxypentos-4-ulose and 3′-Phosphoglycolate Pathways of 4′ Oxidation of 2-Deoxyribose in DNA: Application to DNA Damage Produced by γ Radiation and Bleomycin journal November 2007
53BP1-dependent robust localized KAP-1 phosphorylation is essential for heterochromatic DNA double-strand break repair journal January 2010
Functional and biochemical dissection of the structure-specific nuclease ARTEMIS journal April 2004
Interplay between Ku, Artemis, and the DNA-dependent Protein Kinase Catalytic Subunit at DNA Ends journal September 2006
Processing of 3′-Phosphoglycolate-terminated DNA Double Strand Breaks by Artemis Nuclease journal February 2007
Cytotoxicity Associated with Artemis Overexpression After Lentiviral Vector-Mediated Gene Transfer journal July 2010
Deficiency in 3'-phosphoglycolate processing in human cells with a hereditary mutation in tyrosyl-DNA phosphodiesterase (TDP1) journal January 2005
Coordinate 5′ and 3′ endonucleolytic trimming of terminally blocked blunt DNA double-strand break ends by Artemis nuclease and DNA-dependent protein kinase journal April 2008
 -H2AX in recognition and signaling of DNA double-strand breaks in the context of chromatin journal August 2008
PML Nuclear Bodies journal April 2010
Dynamics of DNA Double-Strand Breaks Revealed by Clustering of Damaged Chromosome Domains journal January 2004
The Ku80 Carboxy Terminus Stimulates Joining and Artemis-Mediated Processing of DNA Ends journal December 2008
Artemis Links ATM to G2/M Checkpoint Recovery via Regulation of Cdk1-Cyclin B journal January 2007
ARTEMIS Nuclease Facilitates Apoptotic Chromatin Cleavage journal October 2009
Nucleosome resection at a double-strand break during Non-Homologous Ends Joining in mammalian cells - implications from repressive chromatin organization and the role of ARTEMIS journal January 2011
A Versatile Viral System for Expression and Depletion of Proteins in Mammalian Cells journal August 2009
Comparison of Repair of DNA Double-Strand Breaks in Identical Sequences in Primary Human Fibroblast and Immortal Hamster–Human Hybrid Cells Harboring a Single Copy of Human Chromosome 11 journal June 2000
gamma Ray induced deoxyribonucleic acid strand breaks. 3' Glycolate termini. journal January 1983
Sequence-specific double-strand breakage of DNA by neocarzinostatin involves different chemical mechanisms within a staggered cleavage site. journal September 1990
Artemis, a Novel DNA Double-Strand Break Repair/V(D)J Recombination Protein, Is Mutated in Human Severe Combined Immune Deficiency journal April 2001
Hairpin Opening and Overhang Processing by an Artemis/DNA-Dependent Protein Kinase Complex in Nonhomologous End Joining and V(D)J Recombination journal March 2002
Artemis deficiency confers a DNA double-strand break repair defect and Artemis phosphorylation status is altered by DNA damage and cell cycle progression journal May 2005
Induction and Rejoining of DNA Double Strand Breaks Assessed by H2AX Phosphorylation in Melanoma Cells Irradiated with Proton and Lithium Beams journal July 2009
A Pathway of Double-Strand Break Rejoining Dependent upon ATM, Artemis, and Proteins Locating to γ-H2AX Foci journal December 2004
ATM Signaling Facilitates Repair of DNA Double-Strand Breaks Associated with Heterochromatin journal July 2008
ATM and Artemis promote homologous recombination of radiation-induced DNA double-strand breaks in G2 journal September 2009
DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation journal January 2003
PML-dependent apoptosis after DNA damage is regulated by the checkpoint kinase hCds1/Chk2 journal October 2002
DNA-PK autophosphorylation facilitates Artemis endonuclease activity journal July 2006
PML NBs associate with the hMre11 complex and p53 at sites of irradiation induced DNA damage journal March 2002
Artemis Regulates Cell Cycle Recovery from the S Phase Checkpoint by Promoting Degradation of Cyclin E journal July 2009
Interplay between Ku, Artemis, and the DNA-dependent Protein Kinase Catalytic Subunit at DNA Ends journal September 2006
Processing of 3′-Phosphoglycolate-terminated DNA Double Strand Breaks by Artemis Nuclease journal February 2007
Promyelocytic Leukemia Protein (Pml) and Daxx Participate in a Novel Nuclear Pathway for Apoptosis journal February 2000
The Ku80 Carboxy Terminus Stimulates Joining and Artemis-Mediated Processing of DNA Ends journal December 2008
Artemis Links ATM to G2/M Checkpoint Recovery via Regulation of Cdk1-Cyclin B journal January 2007
ARTEMIS Nuclease Facilitates Apoptotic Chromatin Cleavage journal October 2009
A Mitotic Phosphorylation Feedback Network Connects Cdk1, Plk1, 53BP1, and Chk2 to Inactivate the G2/M DNA Damage Checkpoint journal January 2010
Comparison of Repair of DNA Double-Strand Breaks in Identical Sequences in Primary Human Fibroblast and Immortal Hamster–Human Hybrid Cells Harboring a Single Copy of Human Chromosome 11 journal June 2000
Hypertonic Saline Enhances Expression of Phosphorylated Histone H2AX after Irradiation journal April 2004
An Imperfect G 2 M Checkpoint Contributes to Chromosome Instability Following Irradiation of S and G 2 Phase Cells journal July 2007

Cited By (8)

Elevated expression of artemis in human fibroblast cells is associated with cellular radiosensitivity and increased apoptosis journal October 2012
Structure-Specific nuclease activities of Artemis and the Artemis: DNA-PKcs complex journal May 2016
TDP1 suppresses mis-joining of radiomimetic DNA double-strand breaks and cooperates with Artemis to promote optimal nonhomologous end joining journal August 2018
The Interaction of the Metallo-Glycopeptide Anti-Tumour Drug Bleomycin with DNA journal May 2018
Processing of Damaged DNA Ends for Double-Strand Break Repair in Mammalian Cells journal January 2012
Involvement of p53 in the Repair of DNA Double Strand Breaks: Multifaceted Roles of p53 in Homologous Recombination Repair (HRR) and Non-Homologous End Joining (NHEJ) book January 2014
Modeling the interplay between DNA-PK, Artemis, and ATM in non-homologous end-joining repair in G1 phase of the cell cycle journal February 2019
Effects of hyperthermia on DNA repair pathways: one treatment to inhibit them all journal August 2015

Similar Records

Coordinateendonucleolytic 5' and 3' trimming of terminally blocked blunt DNA double-strand break ends by Artemis nuclease and DNA-dependent protein kinase
Journal Article · Sun Feb 17 23:00:00 EST 2008 · Nucleic Acids Research · OSTI ID:934731
DNA-PK inhibition causes a low level of H2AX phosphorylation and homologous recombination repair in Medaka (Oryzias latipes) cells
Journal Article · Thu Dec 13 23:00:00 EST 2012 · Biochemical and Biophysical Research Communications · OSTI ID:22210356
Processing of 3'-Phosphoglycolate-Terminated DNA Double-StrandBreaks by Artemis Nuclease
Journal Article · Sat Oct 01 00:00:00 EDT 2005 · Journal of Biological Chemistry · OSTI ID:923183

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Biochemistry & Molecular Biology