In Vivo Analysis of Protein Kinase B (PKB)/Akt Regulation in DNA-PKcs-null Mice Reveals a Role for PKB/Akt in DNA Damage Response and Tumorigenesis

Surucu, Banu; Bozulic, Lana; Hynx, Debby; Parcellier, Arnaud; Hemmings, Brian A.

doi:10.1074/jbc.m803053200

Cited by 66 publications

(53 citation statements)

References 78 publications

(55 reference statements)

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“…Treatment of HCT116 cells with DNA-damaging reagent doxorubicin induced p53 accumulation, with a concomitant increase in Ser473 phosphorylation of PKB (Fig. 3A), consistent with previous studies [20,32].…”

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confidence: 79%

PKB-mediated PHF20 phosphorylation on Ser291 is required for p53 function in DNA damage

Park

et al. 2013

Cellular Signalling

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Publisher rights This is the author's version of a work that was accepted for publication in Cellular Signalling. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Cellular Signalling, VOL25, ISSUE1, 01/2013General rights Copyright for the publications made accessible via the Queen's University Belfast Research Portal is retained by the author(s) and / or other copyright owners and it is a condition of accessing these publications that users recognise and abide by the legal requirements associated with these rights. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Running Head: Regulation of PHF20 by PKB Keywords : PHF20, Glioblastoma, PKB, p53, protein phosphorylation A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT2 Abstract PHD finger protein 20 (PHF20) is a transcription factor, which was originally identified in glioma patients. PHF20 appears to be a novel antigen in glioma, and has also termed gliomaexpressed antigen 2. PHF20 is thought to contribute to the development of cancers, including glioblastoma, lung cancer, colon cancer and ovarian cancer. However, little is known about the function of PHF20 in various cancers. Here we report that PHF20 contains two consensus sites for protein kinase B (PKB) phosphorylation (RxRxxS/T). PKB can directly phosphorylate PHF20 on Ser291 in vitro and in vivo. It has been shown that PKB participates in the tumor suppressor p53 regulated gene expression program and has a direct effect on p21 regulation after DNA damage. UV induced DNA damage result in accumulation of p53 and PKB activation. Interestingly, PKB-mediated PHF20 phosphorylation led to an inhibition of p53 induction following UV treatment, leading to the reduction of p21 transcriptional activity.Using anti PHF20 and anti pPKB (S473) antibodies, these events were mapped in various human cancer tissues. Taken together, these data suggest that PHF20 is a novel substrate for PKB and its phosphorylation by PKB plays an important role in tumorigenesis via regulating of p53 mediated signaling.

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confidence: 79%

PKB-mediated PHF20 phosphorylation on Ser291 is required for p53 function in DNA damage

Park

et al. 2013

Cellular Signalling

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“…4B). In addition, there is a much higher degree of AKT1 S473 phosphorylation in DNA-PKcs wild-type cells, in accordance with reports, that S473 is a target of DNA-PK (33,34). Phosphorylation of GSK3β (S9), a downstream target of AKT1, mirrored the AKT1 S473 phosphorylation in DNA-PKcs expressing cells with respect to NCS treatment and also with respect to TBB (Fig.…”

Section: Resultssupporting

confidence: 75%

“…Homozygous BID knockout mice display chromosomal abnormalities and develop a disorder resembling chronic myelomonocytic leukemia (30,31), which indicates that BID plays a role in the DNA damage response and in genomic stability. AKT1 is activated following DSBs through phosphorylation by DNA-PK and PDK1 at Ser473 and Thr308, respectively (32)(33)(34). AKT1 knockout mice show impaired DNA damage-dependent induction of the cyclindependent kinase inhibitor p21 and increased apoptosis (34), indicating that AKT1 plays a role in promoting survival in response to DNA damage.…”

Section: Introductionmentioning

confidence: 99%

Characterization of ATM and DNA-PK wild-type and mutant cell lines upon DSB induction in the presence and absence of CK2 inhibitors

2011

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Abstract. Protein kinase CK2 is involved in several cellular processes and has lately also been linked to the DNA damage response through phosphorylations and interactions. Herein, we have analysed two sets of mouse cell lines, one pair, which is proficient and deficient in ATM and the other set expressing or lacking a functional catalytic subunit of the DNA dependent protein kinase (DNA-PKcs). Both kinases are implicated in the downstream phosphorylation of the signaling molecules such as BID and AKT1 in response to DNA damage. BID and AKT1 are also targets of CK2, hence the four cell lines were treated with the three established CK2 inhibitors DMAT, TBB and resorufin in the absence and presence of the radiomimetic drug neocarzinostatin, which induces DNA double-strand breaks. We show that there are differences with respect to the effect of the CK2 inhibitors on the phosphorylation of AKT1 at S473 and its downstream target GSK3β as well as between the two sets of cell lines. However, no such change was seen with BID phosphorylation. The most notably difference was the higher expression of CK2α' and CK2β in DNA-PKcs defective cells compared to the DNA-PKcs proficient cells.

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“…Consistent with previous reports (39,40), treatment of cells with doxorubicin, a DNA damage agent, increased Ser-473 phosphorylation of PKB in DNA-PK ϩ/ϩ MEFs, but not DNA-PK Ϫ/Ϫ MEFs (31, 32), indicating that DNA-PK is required Ser-473 phosphorylation and that PKB activation also occurred during DNA repair signaling. We have previously shown that insulin-stimulated PKB phosphorylation on Ser-473 did not occur through DNA-PK and that ␥-irradiation triggered DNA-PK mediated phosphorylation on this residue (32). We now extend these findings to define the amino acids in the hydrophobic motif of PKB required for DNA-PK mediated Ser-473 phosphorylation.…”

Section: Discussionmentioning

confidence: 55%

“…It was previously shown that PKB␣ knock-out mice show impaired DNA damage-dependent induction of p21 after single dose whole body irradiation (31). In addition, phosphorylation of the PKB target FoxO4 on Ser-193, which increases in doxyrubicin-treated cells, was abolished in DNA-PK Ϫ/Ϫ cells (32). Therefore, two events (Ser-193 phosphorylation of FoxO4 and p21 protein levels) were monitored in cells expressing PKB mutants following doxorubicin treatment.…”

Section: Dna-pk Effectively Phosphorylates Fsytide Whenmentioning

confidence: 99%

DNA-dependent Protein Kinase-mediated Phosphorylation of Protein Kinase B Requires a Specific Recognition Sequence in the C-terminal Hydrophobic Motif

Park

Feng

et al. 2009

Journal of Biological Chemistry

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DNA-dependent protein kinase (DNA-PK) has been implicated in a variety of nuclear processes including DNA double strand break repair, V(D)J recombination, and transcription. A recent study showed that DNA-PK is responsible for Ser-473 phosphorylation in the hydrophobic motif of protein kinase B (PKB/Akt) in genotoxic-stressed cells, suggesting a novel role for DNA-PK in cell signaling. Here, we report that DNA-PK activity toward PKB peptides is impaired in DNA-PK knock-out mouse embryonic fibroblast cells when compared with wild type. In addition, human glioblastoma cells expressing a mutant form of DNA-PK (M059J) displayed a lower DNA-PK activity when compared with glioblastoma cells expressing wild-type DNA-PK (M059K) when PKB peptide substrates were tested. DNA-PK preferentially phosphorylated PKB on Ser-473 when compared with its known in vitro substrate, p53. A consensus hydrophobic amino acid surrounding the Ser-473 phospho-acceptor site in PKB containing amino acids Phe at position ؉1 and ؉4 and Tyr at position ؊1 are critical for DNA-PK activity. Thus, these data define the specificity of DNA-PK action as a Ser-473 kinase for PKB in DNA repair signaling.

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In Vivo Analysis of Protein Kinase B (PKB)/Akt Regulation in DNA-PKcs-null Mice Reveals a Role for PKB/Akt in DNA Damage Response and Tumorigenesis

Cited by 66 publications

References 78 publications

PKB-mediated PHF20 phosphorylation on Ser291 is required for p53 function in DNA damage

PKB-mediated PHF20 phosphorylation on Ser291 is required for p53 function in DNA damage

Characterization of ATM and DNA-PK wild-type and mutant cell lines upon DSB induction in the presence and absence of CK2 inhibitors

DNA-dependent Protein Kinase-mediated Phosphorylation of Protein Kinase B Requires a Specific Recognition Sequence in the C-terminal Hydrophobic Motif

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