Cited by 144DNA Repair MechanismsGenomics and Chromatin DynamicsCRISPR and Genetic Engineering

The DNA-dependent protein kinase catalytic subunit phosphorylation sites in human Artemis

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Ma, Yunmei, Pannicke, Ulrich, Lu, Haihui, Niewolik, Doris, Schwarz, Klaus, Lieber, Michael R.·2005·Journal of Biological Chemistry

Abstract

Artemis protein has irreplaceable functions in V(D)J recombination and nonhomologous end joining (NHEJ) as a hairpin and 5' and 3' overhang endonuclease. The kinase activity of the DNA-dependent protein kinase catalytic subunit (DNA-PKcs) is necessary in activating Artemis as an endonuclease. Here we report that three basal phosphorylation sites and 11 DNA-PKcs phosphorylation sites within the mammalian Artemis are all located in the C-terminal domain. All but one of these phosphorylation sites deviate from the SQ or TQ motif of DNA-PKcs that was predicted previously from in vitro phosphorylation studies. Phosphatase-treated mammalian Artemis and Artemis that is mutated at the three basal phosphorylation sites still retain DNA-PKcs-dependent endonucleolytic activities, indicating that basal phosphorylation is not required for the activation. In vivo studies of Artemis lacking the C-terminal domain have been reported to be sufficient to complement V(D)J recombination in Artemis null cells. Therefore, the C-terminal domain may have a negative regulatory effect on the Artemis endonucleolytic activities, and phosphorylation by DNA-PKcs in the C-terminal domain may relieve this inhibition.

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Authors
Ma, Y., Pannicke, U., Lu, H., Niewolik, D., Schwarz, K., & Lieber, M. R.
Affiliations
  • University of Southern California (US)
  • Universität Ulm (DE)
Corresponding author
  • Schwarz, Klaus
  • Lieber, Michael R.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2005-10
Volume
280
Issue
40
Pages
33839-33846
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2088698677
Cited by
144
References
32
Citations by year
2026: 1 · 2024: 3 · 2023: 5 · 2022: 6 · 2021: 2 · 2020: 3 · 2019: 3 · 2018: 6 · 2017: 6 · 2016: 2 · 2015: 3 · 2014: 4 · 2013: 11 · 2012: 9
FWCI
4.14
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
University of Virginia
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
Keywords
Artemis, DNA-PKcs, phosphorylation sites, C-terminal domain, V(D)J recombination, non-homologous end joining, endonuclease activation, phosphatase treatment
MeSH terms
DNA, DNA-Binding Proteins, Endonucleases, Humans, Nuclear Proteins, Phosphorylation, Plasmids, Recombination, Genetic, Polymerase Chain Reaction, Mutagenesis, Site-Directed, Baculoviridae, Catalytic Domain, DNA-Activated Protein Kinase

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