The N-terminal Region of the DNA-dependent Protein Kinase Catalytic Subunit Is Required for Its DNA Double-stranded Break-mediated Activation

Davis, Anthony J.; Lee, Kyung Jong; Chen, David J.

doi:10.1074/jbc.m112.434498

Cited by 35 publications

(30 citation statements)

References 57 publications

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“…Contrary to the paradigm that binding to DNA is a requirement for DNA-PKcs activation, it was demonstrated that DNA-PKcs autophosphorylation can be modulated by N-terminal conformational changes in the protein (such as that likely caused by DNA binding) [52]. This supposition was further supported by findings which revealed that the N-terminus (but not the C-terminus) is required for binding to the Ku-DNA complex and robust DNA-PKcs activity [53]. These collective studies support the concept that the N-terminus of DNA-PKcs is crucial for both targeting to Ku-bound DNA, and for conformational change-induced enzymatic activity.…”

Section: Regulation Of Dna-pk Activitymentioning

confidence: 99%

Beyond DNA Repair: DNA-PK Function in Cancer

2014

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The DNA-dependent protein kinase (DNA-PK) is a pivotal component of the DNA repair machinery that governs the response to DNA damage, serving to maintain genome integrity. However, the DNA-PK kinase component was initially isolated with transcriptional complexes, and recent findings have illuminated the impact of DNA-PK-mediated transcriptional regulation on tumor progression and therapeutic response. DNA-PK expression has also been correlated with poor outcome in selected tumor types, further underscoring the importance of understanding its role in disease. Herein, the molecular and cellular consequence of DNA-PK will be considered, with an eye toward discerning the rationale for therapeutic targeting of DNA-PK.

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Section: Regulation Of Dna-pk Activitymentioning

confidence: 99%

Beyond DNA Repair: DNA-PK Function in Cancer

2014

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“…DNA double-strand breaks (DSBs) 1 occur frequently during cell cycle progression and are corrected either by non-homologous end joining (NHEJ) or homologous recombination (HR). DNA-dependent protein kinase (DNA-PK) plays a key role in NHEJ-mediated repair of DSBs, and is comprised of a DNAbinding heterodimer Ku70/Ku80 and a catalytic subunit (DNAPKcs) that are necessary for the kinase activity of the enzyme (45). Euchromatin DSBs are repaired by NHEJ during G 1 and G 2 phase, whereas heterochromatin DSB repair is mediated by HR during G 2 phase (46,47), and perhaps also during S phase (48).…”

Section: Mass Spectrometric Identification and Quantification Of Thementioning

confidence: 99%

Profiling of the Chromatin-associated Proteome Identifies HP1BP3 as a Novel Regulator of Cell Cycle Progression

Dutta

Ren

Hao

et al. 2014

Molecular & Cellular Proteomics

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The chromatin-associated proteome (chromatome) regulates cellular gene expression by restricting access of transcriptional machinery to template DNA, and dynamic re-modeling of chromatin structure is required to regulate critical cell functions including growth and replication, DNA repair and recombination, and oncogenic transformation in progression to cancer. Central to the control of these processes is efficient regulation of the host cell cycle, which is maintained by rapid changes in chromatin conformation during normal cycle progression. A global overview of chromatin protein organization is therefore essential to fully understand cell cycle regulation, but the influence of the chromatome and chromatin binding topology on host cell cycle progression remains poorly defined. Here we used partial MNase digestion together with iTRAQ-based high-throughput quantitative proteomics to quantify chromatin-associated proteins during interphase progression. We identified a total of 481 proteins with high confidence that were involved in chromatin-dependent events including transcriptional regulation, chromatin reorganization, and DNA replication and repair, whereas the quantitative data revealed the temporal interactions of these proteins with chromatin during interphase progression. When combined with biochemical and functional assays, these data revealed a strikingly dynamic association of protein HP1BP3 with the chromatin complex during different stages of interphase, and uncovered a novel regulatory role for this molecule in transcriptional regulation. We report that HP1BP3 protein maintains heterochromatin integrity during G 1-S progression and regulates the duration of G 1 phase to critically influence cell proliferative capacity. Molecular & Cellular

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“…DNA-PKcs is composed of HEAT (Huntington-elongation-A-subunit-TOR) repeats in its N-terminus, which produce a pincer-shaped structure that forms a central channel and a C-terminal region that contains the PI3 kinase domain, which is flanked by the FAT (FRAP, ATM, TRRAP) domain at its N-terminal side and by the FATC domain at its C-terminal side [5, 6]. Following DSB induction, the Ku heterodimer quickly binds to the DSB ends and recruits DNA-PKcs to the break site, which is mediated by the N-terminal region of DNA-PKcs [7–9]. Upon interacting with the DSB-Ku complex, DNA-PKcs is activated.…”

Section: Introductionmentioning

confidence: 99%

Lysines 3241 and 3260 of DNA-PKcs are important for genomic stability and radioresistance

Mori

Davis

Hasegawa

et al. 2016

Biochemical and Biophysical Research Communications

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DNA-dependent protein kinase (DNA-PK) is a serine/threonine kinase that plays an essential role in the repair of DNA double-strand breaks (DSBs) in the non-homologous end-joining (NHEJ) pathway. The DNA-PK holoenzyme consists of a catalytic subunit (DNA-PKcs) and DNA-binding subunit (Ku70/80, Ku). Ku is a molecular sensor for double-stranded DNA and once bound to DSB ends it recruits DNA-PKcs to the DSB site. Subsequently, DNA-PKcs is activated and heavily phosphorylated, with these phosphorylations modulating DNA-PKcs. Although phosphorylation of DNA-PKcs is well studied, other post-translational modifications of DNA-PKcs are not. In this study, we aimed to determine if acetylation of DNA-PKcs regulates DNA-PKcs-dependent DSB repair. We report that DNA-PKcs is acetylated in vivo and identified two putative acetylation sites, lysine residues K3241 and K3260. Mutating these sites to block potential acetylation results increased radiosensitive, a slight decrease in DSB repair capacity as assessed by γH2AX resolution, and increased chromosomal aberrations, especially quadriradial chromosomes. Together, our results provide evidence that acetylation potentially regulates DNA-PKcs.

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The N-terminal Region of the DNA-dependent Protein Kinase Catalytic Subunit Is Required for Its DNA Double-stranded Break-mediated Activation

Cited by 35 publications

References 57 publications

Beyond DNA Repair: DNA-PK Function in Cancer

Beyond DNA Repair: DNA-PK Function in Cancer

Profiling of the Chromatin-associated Proteome Identifies HP1BP3 as a Novel Regulator of Cell Cycle Progression

Lysines 3241 and 3260 of DNA-PKcs are important for genomic stability and radioresistance

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