7TYR

Cryo-EM structure of the basal state of the Artemis:DNA-PKcs complex (see COMPND 13/14)

Summary for 7TYR

• Entry DOI: 10.2210/pdb7tyr/pdb
• EMDB information: 26192
• Descriptor: DNA-dependent protein kinase catalytic subunit, Protein artemis (2 entities in total)
• Functional Keywords: kinase, nuclease, dna binding protein
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 2
• Total formula weight: 550166.57

Authors

Watanabe, G.,Lieber, M.R.,Williams, D.R. (deposition date: 2022-02-14, release date: 2022-07-20, Last modification date: 2024-11-13)

Primary citation

Watanabe, G.,Lieber, M.R.,Williams, D.R.
Structural analysis of the basal state of the Artemis:DNA-PKcs complex.
Nucleic Acids Res., 50:7697-7720, 2022

PubMed Abstract: Artemis nuclease and DNA-dependent protein kinase catalytic subunit (DNA-PKcs) are key components in nonhomologous DNA end joining (NHEJ), the major repair mechanism for double-strand DNA breaks. Artemis activation by DNA-PKcs resolves hairpin DNA ends formed during V(D)J recombination. Artemis deficiency disrupts development of adaptive immunity and leads to radiosensitive T- B- severe combined immunodeficiency (RS-SCID). An activated state of Artemis in complex with DNA-PK was solved by cryo-EM recently, which showed Artemis bound to the DNA. Here, we report that the pre-activated form (basal state) of the Artemis:DNA-PKcs complex is stable on an agarose-acrylamide gel system, and suitable for cryo-EM structural analysis. Structures show that the Artemis catalytic domain is dynamically positioned externally to DNA-PKcs prior to ABCDE autophosphorylation and show how both the catalytic and regulatory domains of Artemis interact with the N-HEAT and FAT domains of DNA-PKcs. We define a mutually exclusive binding site for Artemis and XRCC4 on DNA-PKcs and show that an XRCC4 peptide disrupts the Artemis:DNA-PKcs complex. All of the findings are useful in explaining how a hypomorphic L3062R missense mutation of DNA-PKcs could lead to insufficient Artemis activation, hence RS-SCID. Our results provide various target site candidates to design disruptors for Artemis:DNA-PKcs complex formation.

PubMed: 35801871
DOI: 10.1093/nar/gkac564

Experimental method

ELECTRON MICROSCOPY (3.33 Å)