DNA-PKcs and PIDD interaction in DNA damage response

DNA 损伤反应中 DNA-PKcs 和 PIDD 相互作用

基本信息

批准号：
10693192
负责人：
Anthony J Davis
金额：
$ 36.32万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10693192
关键词：
Acceleration Accidents Affect Apoptosis Attenuated B-Cell Lymphomas CASP2 gene Cell Culture Techniques Cell Death Cell Differentiation process Cell Line Cell Proliferation Cell Survival Cells Complex DNA DNA Damage DNA Double Strand Break DNA Repair DNA Repair Pathway DNA lesion DNA replication fork DNA-PKcs DNA-dependent protein kinase Death Domain Development Double Strand Break Repair G22P1 gene Genome Holoenzymes In Vitro Incidence Inflammation Investigation Ionizing radiation Liver Malignant Neoplasms Malignant neoplasm of liver Mediating Mediator Mus Mutation Nonhomologous DNA End Joining Normal Cell Oncogene Activation Oncogenes Pathway interactions Phosphorylation Phosphotransferases Play Primary carcinoma of the liver cells Process Proteins Radiation Tolerance Radiation exposure Reagent Recovery Regulation Reporting Resistance Role Signal Pathway Steatohepatitis TP53 gene Testing Tissues Ultraviolet Rays Work ataxia telangiectasia mutated protein biological adaptation to stress cancer cell cancer prevention carcinogenesis genome integrity in vivo liver cancer model male mouse model mutant novel preservation prevent recruit regenerative repaired replication stress response senescence tumorigenesis ultraviolet irradiation

项目摘要

Project Summary/Abstract The DNA dependent protein kinase catalytic subunit (DNA-PKcs) is a key regulator of the canonical non- homologous end-joining (NHEJ) pathway for repair of DNA double strand breaks (DSBs) and resistance to ionizing radiation (IR). DNA-PKcs is recruited by the Ku70/80 heterodimer to the DSB ends to form the DNA-PK holoenzyme to initiate NHEJ mechanism. DNA-PKcs also plays an important role in cellular resistance to replication stress. It coordinates with and is rapidly phosphorylated by the ATR (ataxia telangiectasia mutated and Rad3 related) kinase at stalled replication forks upon UV, although the mechanism is not well understood. Our recent work has focused on identifying the player(s) required for the recruitment of DNA-PKcs to stalled replication forks. We have identified that PIDD (p53-induced protein with a death domain), a known apoptosis mediator for assembling the PIDDosome complex and Caspase-2 activation, is required for DNA-PKcs recruitment to stalled replication forks and its association with ATR. Disrupting the interaction between DNA- PKcs and PIDD not only compromised ATR dependent DNA-PKcs phosphorylation at the Thr2609 cluster but also attenuated the ATR signaling pathway and intra-S checkpoint. To assist our investigation, we have created cell lines and a mouse model expressing a DNA-PKcsPL mutant protein unable to interact with PIDD. Our results showed that DNA-PKcsPL cells were highly sensitive to both UV and IR. Based on these preliminary findings, we hypothesize that PIDD, but not the Ku70/80 heterodimer, mediates DNA-PKcs recruitment to stalled replication forks and promotes its association with the ATR pathway. We also hypothesize that PIDD facilitates DNA-PKcs kinase activation and DSB repair. Finally, we hypothesize that the interaction of DNA-PKcs with PIDD will affect cell death regulation and cancer development. In this project, we will determine the coordination between DNA- PKcs, PIDD, and ATR in the cellular response to replication stress. Our specific aims are: 1) To test the hypothesis that PIDD modulates DNA-PKcs kinase activation at stalled replication forks and at IR-induced DNA lesions that are not bound by Ku70/80, 2) To test the hypothesis that PIDD is required for DNA-PKcs to properly function at stalled replication forks upon UV irradiation, and 3) To test the hypothesis that the DNA-PKcs-PIDD association affects cell fate determination upon DNA damage and cancer development.

项目摘要/摘要 DNA依赖性蛋白激酶催化亚基（DNA-PKC）是规范非 - 的关键调节剂 DNA双链断裂（DSB）修复的同源最终连接（NHEJ）途径电离辐射（IR）。 DNA-PKC由KU70/80异二聚体募集到DSB末端以形成DNA-PK 全酶启动NHEJ机制。 DNA-PKC在细胞阻力中也起着重要作用复制应力。它与ATR（共济失调突变的ata省尽管该机理尚不清楚，但在紫外线上的复制叉处的RAD3相关）激酶。我们最近的工作重点是确定招募DNA-PKC所需的玩家复制叉。我们已经确定PIDD（p53诱导的具有死亡结构域的蛋白），一种已知的凋亡 DNA-PKC需要组装Piddosome复合物和caspase-2激活的介体招募停滞的复制叉及其与ATR的关联。破坏DNA-之间的相互作用 PKC和PIDD不仅损害了Thr2609群集的ATR依赖性DNA-PKCS磷酸化，还会损害还减弱了ATR信号通路和内部检查点。为了协助我们的调查，我们创建了表达DNA-PKCSPL突变体蛋白无法与PIDD相互作用的细胞系和小鼠模型。我们的结果表明DNA-PKCSPL细胞对UV和IR都高度敏感。根据这些初步发现，我们假设PIDD，但不是KU70/80异二聚体，将DNA-PKCS募集介导了降低的复制叉子并促进其与ATR途径的关联。我们还假设PIDD促进DNA-PKCS 激酶激活和DSB修复。最后，我们假设DNA-PKC与PIDD的相互作用会影响细胞死亡调节和癌症发展。在这个项目中，我们将确定DNA-之间的协调 PKC，PIDD和ATR在对复制应力的细胞反应中。我们的具体目的是：1）测试 PIDD在停滞的复制叉和IR诱导的DNA处调节DNA-PKCS激酶激活的假设不受KU70/80的约束的病变，以检验DNA-PKC需要PIDD正确的假设紫外线照射时停滞的复制叉的功能，以及3）测试DNA-PKCS-PIDD的假设关联会影响细胞命运对DNA损伤和癌症发展。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

DNA-PKcs and ATM modulate mitochondrial ADP-ATP exchange as an oxidative stress checkpoint mechanism.

DOI：
10.15252/embj.2022112094
发表时间：
2023-03-15
期刊：
The EMBO journal
影响因子：
0
作者：
通讯作者：

Personalized Ultrafractionated Stereotactic Adaptive Radiotherapy (PULSAR) in Preclinical Models Enhances Single-Agent Immune Checkpoint Blockade.

DOI：
10.1016/j.ijrobp.2021.03.047
发表时间：
2021-08-01
期刊：
International journal of radiation oncology, biology, physics
影响因子：
0
作者：
Moore C;Hsu CC;Chen WM;Chen BPC;Han C;Story M;Aguilera T;Pop LM;Hannan R;Fu YX;Saha D;Timmerman R
通讯作者：
Timmerman R

Lysophosphatidic Acid Receptor 3 Promotes Mitochondrial Homeostasis against Oxidative Stress: Potential Therapeutic Approaches for Hutchinson-Gilford Progeria Syndrome.