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TitleDimers of DNA-PK create a stage for DNA double-strand break repair.
Journal, issue, pagesNat Struct Mol Biol, Vol. 28, Issue 1, Page 13-19, Year 2021
Publish dateOct 19, 2020
AuthorsAmanda K Chaplin / Steven W Hardwick / Shikang Liang / Antonia Kefala Stavridi / Ales Hnizda / Lee R Cooper / Taiana Maia De Oliveira / Dimitri Y Chirgadze / Tom L Blundell /
PubMed AbstractDNA double-strand breaks are the most dangerous type of DNA damage and, if not repaired correctly, can lead to cancer. In humans, Ku70/80 recognizes DNA broken ends and recruits the DNA-dependent ...DNA double-strand breaks are the most dangerous type of DNA damage and, if not repaired correctly, can lead to cancer. In humans, Ku70/80 recognizes DNA broken ends and recruits the DNA-dependent protein kinase catalytic subunit (DNA-PKcs) to form DNA-dependent protein kinase holoenzyme (DNA-PK) in the process of non-homologous end joining (NHEJ). We present a 2.8-Å-resolution cryo-EM structure of DNA-PKcs, allowing precise amino acid sequence registration in regions uninterpreted in previous 4.3-Å X-ray maps. We also report a cryo-EM structure of DNA-PK at 3.5-Å resolution and reveal a dimer mediated by the Ku80 C terminus. Central to dimer formation is a domain swap of the conserved C-terminal helix of Ku80. Our results suggest a new mechanism for NHEJ utilizing a DNA-PK dimer to bring broken DNA ends together. Furthermore, drug inhibition of NHEJ in combination with chemo- and radiotherapy has proved successful, making these models central to structure-based drug targeting efforts.
External linksNat Struct Mol Biol / PubMed:33077952
MethodsEM (single particle)
Resolution3.24 - 7.24 Å
Structure data

11185

6zfp

EMDB-11185, PDB-6zfp:
Cryo-EM structure of DNA-PKcs (State 2)
Method: EM (single particle) / Resolution: 3.24 Å

11211

6zh2

EMDB-11211, PDB-6zh2:
Cryo-EM structure of DNA-PKcs (State 1)
Method: EM (single particle) / Resolution: 3.92 Å

11213

6zh4

EMDB-11213, PDB-6zh4:
Cryo-EM structure of DNA-PKcs (State 3)
Method: EM (single particle) / Resolution: 3.62 Å

11215

6zh6

EMDB-11215, PDB-6zh6:
Cryo-EM structure of DNA-PKcs:Ku80ct194
Method: EM (single particle) / Resolution: 3.93 Å

11216

6zh8

EMDB-11216, PDB-6zh8:
Cryo-EM structure of DNA-PKcs:DNA
Method: EM (single particle) / Resolution: 4.14 Å

11217

6zha

EMDB-11217, PDB-6zha:
Cryo-EM structure of DNA-PK monomer
Method: EM (single particle) / Resolution: 3.91 Å

11219

6zhe

EMDB-11219, PDB-6zhe:
Cryo-EM structure of DNA-PK dimer
Method: EM (single particle) / Resolution: 7.24 Å

Source
  • homo sapiens (human)
KeywordsDNA BINDING PROTEIN / Kinase / DNA-PKcs / NHEJ / DNA-repair / DNA-PK / Ku80 / Ku70/80

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