Structure of an activated DNA-PK and its implications for NHEJ

Chen, Xuemin; Xu, Xiang; Chen, Yun; Cheung, Joyce; Wang, Huaibin; Jiang, Junchen; Val, Natalia de; Fox, Tara; Gellert, Martin; Yang, Wei

doi:10.1016/j.molcel.2020.12.015

Cited by 100 publications

(158 citation statements)

References 61 publications

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“…Since 2017, members of the Blundell research group reported crystals of selenomethionine (Se-Met)-labeled DNA-PKcs complexed with native C-terminal of Ku80 (residues 539-732) diffracting to 4.3 Å resolution (Sibanda et al, 2017). Furthermore, several cryo-EM structures have been reported of the apo DNA-PKcs at 4.8 Å (Sharif et al, 2017), DNA-PK holoenzyme at 6.6 Å (Yin et al, 2017), apo DNA-PKcs at 2.8 Å and dimeric DNA-PK at 7.2 Å (Chaplin et al, 2021), and DNA-PK in a series of different states up to 3.7 Å (Chen et al, 2021b). The data from these studies have revealed how the DNA-PK holoenzyme might be assembled and activated.…”

Section: Dna-pkcsmentioning

confidence: 99%

“…The structural units of DNA-PKcs comprise the N-terminal region (residues 1-892), the circular cradle (residues 893-2801), and the C-terminal head (residues 2802-4128, FAT, FRB, kinase, and FATC) (Fig. 1G) (Chaplin et al, 2021;Chen et al, 2021b;Sharif et al, 2017;Sibanda et al, 2017;Yin et al, 2017). DNA-PKcs alone have weak basal activity (Gottlieb & Jackson, 1993;Hartley et al, 1995;Lees-Miller et al, 1990;Yin et al, 2017).…”

Section: Dna-pkcsmentioning

confidence: 99%

“…The PRD domain lies adjacent to FRB and blocks the active site entrance, suggesting an inactive conformation (Fig. 4A) (Chen et al, 2021b;Sibanda et al, 2017).…”

Section: Dna-pkcsmentioning

confidence: 99%

“…The blocked or Y-shaped end is found outside Ku70/80 and the free DNA end is located within the open cradle (Fig. 4B) where it is protected by DNA-PKcs from unnecessary DNA end processing (Chen et al, 2021b;Yin et al, 2017). The structure of apo DNA-PKcs with a DNA oligomer has been determined: the data suggest that DNA-PKcs alone has weak DNA-end binding affinity (Chen et al, 2021b).…”

Section: Dna-pkcsmentioning

confidence: 99%

“…4B) where it is protected by DNA-PKcs from unnecessary DNA end processing (Chen et al, 2021b;Yin et al, 2017). The structure of apo DNA-PKcs with a DNA oligomer has been determined: the data suggest that DNA-PKcs alone has weak DNA-end binding affinity (Chen et al, 2021b). Ku70/80 is situated outside DNA-PKcs, where it bridges the two sides of the DNA-binding groove in DNA-PKcs and stabilizes the DNA-PKcs-DNA interaction (Chaplin et al, 2021;Chen et al, 2021b;Yin et al, 2017).…”

Section: Dna-pkcsmentioning

confidence: 99%

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The activation mechanisms of master kinases in the DNA damage response

Xiao

Rao

2021

GENOME INSTAB. DIS.

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DNA damage can be introduced by intrinsic or extrinsic stimuli and threaten the transmission of genetic information and ultimately, survival. Eukaryotes have evolved a sophisticated mechanism, DNA damage response (DDR), to counteract this threat posed by DNA damage. The DDR requires a series of proteins involved in multiple DNA repair pathways, of which ATM, ATR and DNA-PK serve as the three most critical kinases. The signaling mechanisms underlying these pathways have been extensively studied and these three master kinases structures determined by cryo-electron microscopy (cryo-EM) have led to tremendous progress in understanding the molecular mechanism of the DDR. In this review, we discuss the recent advances made in understanding the structures of ATM, ATR, and DNA-PK. We highlight the implications of these structures in terms of their function and regulation, and speculate on the critical role of PIKK regulatory domain (PRD) in their activation.

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Section: Dna-pkcsmentioning

confidence: 99%

Section: Dna-pkcsmentioning

confidence: 99%

“…The PRD domain lies adjacent to FRB and blocks the active site entrance, suggesting an inactive conformation (Fig. 4A) (Chen et al, 2021b;Sibanda et al, 2017).…”

Section: Dna-pkcsmentioning

confidence: 99%

Section: Dna-pkcsmentioning

confidence: 99%

Section: Dna-pkcsmentioning

confidence: 99%

See 3 more Smart Citations

The activation mechanisms of master kinases in the DNA damage response

Xiao

Rao

2021

GENOME INSTAB. DIS.

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TRIM24 Cooperates with Ras Mutation to Drive Glioma Progression through snoRNA Recruitment of PHAX and DNA‐PKcs

Xu,

Chen,

et al. 2024

Advanced Science

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The factors driving glioma progression remain poorly understood. Here, the epigenetic regulator TRIM24 is identified as a driver of glioma progression, where TRIM24 overexpression promotes HRasV12 anaplastic astrocytoma (AA) progression into epithelioid GBM (Ep‐GBM)‐like tumors. Co‐transfection of TRIM24 with HRasV12 also induces Ep‐GBM‐like transformation of human neural stem cells (hNSCs) with tumor protein p53 gene (TP53) knockdown. Furthermore, TRIM24 is highly expressed in clinical Ep‐GBM specimens. Using single‐cell RNA‐sequencing (scRNA‐Seq), the authors show that TRIM24 overexpression impacts both intratumoral heterogeneity and the tumor microenvironment. Mechanically, HRasV12 activates phosphorylated adaptor for RNA export (PHAX) and upregulates U3 small nucleolar RNAs (U3 snoRNAs) to recruit Ku‐dependent DNA‐dependent protein kinase catalytic subunit (DNA‐PKcs). Overexpressed TRIM24 is also recruited by PHAX to U3 snoRNAs, thereby facilitating DNA‐PKcs phosphorylation of TRIM24 at S767/768 residues. Phosphorylated TRIM24 induces epigenome and transcription factor network reprogramming and promotes Ep‐GBM‐like transformation. Targeting DNA‐PKcs with the small molecule inhibitor NU7441 synergizes with temozolomide to reduce Ep‐GBM tumorigenicity and prolong animal survival. These findings provide new insights into the epigenetic regulation of Ep‐GBM‐like transformation and suggest a potential therapeutic strategy for patients with Ep‐GBM.

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Against all odds: The road to success in the development of human immune reconstitution mice

Bin,

Ren,

Zhang

et al. 2024

Anim Models and Exp Med

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The mouse genome has a high degree of homology with the human genome, and its physiological, biochemical, and developmental regulation mechanisms are similar to those of humans; therefore, mice are widely used as experimental animals. However, it is undeniable that interspecies differences between humans and mice can lead to experimental errors. The differences in the immune system have become an important factor limiting current immunological research. The application of immunodeficient mice provides a possible solution to these problems. By transplanting human immune cells or tissues, such as peripheral blood mononuclear cells or hematopoietic stem cells, into immunodeficient mice, a human immune system can be reconstituted in the mouse body, and the engrafted immune cells can elicit human‐specific immune responses. Researchers have been actively exploring the development and differentiation conditions of host recipient animals and grafts in order to achieve better immune reconstitution. Through genetic engineering methods, immunodeficient mice can be further modified to provide a favorable developmental and differentiation microenvironment for the grafts. From initially only being able to reconstruct single T lymphocyte lineages, it is now possible to reconstruct lymphoid and myeloid cells, providing important research tools for immunology‐related studies. In this review, we compare the differences in immune systems of humans and mice, describe the development history of human immune reconstitution from the perspectives of immunodeficient mice and grafts, and discuss the latest advances in enhancing the efficiency of human immune cell reconstitution, aiming to provide important references for immunological related researches.

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Structure of an activated DNA-PK and its implications for NHEJ

Cited by 100 publications

References 61 publications

The activation mechanisms of master kinases in the DNA damage response

The activation mechanisms of master kinases in the DNA damage response

TRIM24 Cooperates with Ras Mutation to Drive Glioma Progression through snoRNA Recruitment of PHAX and DNA‐PKcs

Against all odds: The road to success in the development of human immune reconstitution mice

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