دورية أكاديمية
Structural analysis of the basal state of the Artemis:DNA-PKcs complex
| العنوان: | Structural analysis of the basal state of the Artemis:DNA-PKcs complex |
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| المؤلفون: | Watanabe, Go, Lieber, Michael R, Williams, Dewight R |
| المساهمون: | National Institutes of Health |
| المصدر: | Nucleic Acids Research ; volume 50, issue 13, page 7697-7720 ; ISSN 0305-1048 1362-4962 |
| بيانات النشر: | Oxford University Press (OUP) |
| سنة النشر: | 2022 |
| الوصف: | 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. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| DOI: | 10.1093/nar/gkac564 |
| الإتاحة: | https://doi.org/10.1093/nar/gkac564Test https://academic.oup.com/nar/article-pdf/50/13/7697/45034164/gkac564.pdfTest |
| حقوق: | https://creativecommons.org/licenses/by/4.0Test/ |
| رقم الانضمام: | edsbas.1252AE0C |
| قاعدة البيانات: | BASE |
| DOI: | 10.1093/nar/gkac564 |
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