Regulation of genomic stability by ORC

ORC 调节基因组稳定性

• 批准号: 10379725
• 负责人: Supriya G Prasanth
• 金额: $ 12.5万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10379725
• 关键词: Address; Biochemical; Biological; Biomedical Research; Cell Cycle Progression; Cell Survival; Cells; Chromatin; Cytokinesis; DNA Damage; DNA biosynthesis; DNA replication fork; Data; Genome; Genome Stability; Genotoxic Stress; Goals; Heterochromatin; Human; Image; Individual; Laboratories; Maintenance; Malignant Neoplasms; Mission; Mitosis; Molecular; Pathway interactions; Phosphorylation; Play; Positioning Attribute; Post-Translational Protein Processing; Proteins; Public Health; Regulation; Replication Initiation; Research; Resolution; Role; S Phase; Techniques; Time; United States National Institutes of Health; base; biophysical techniques; cancer biomarkers; cell growth; combat; experience; genome integrity; innovation; insight; malignant breast neoplasm; novel; origin recognition complex; oxidative DNA damage; prevent; response; single molecule; therapeutic target; therapeutically effective; tumorigenesis

Maintenance of genomic integrity is paramount to cell survival. Precise duplication of our genome and appropriate cellular response to genotoxic stress is critical to maintain genome stability. Origin Recognition Complex (ORC, composed of six subunits) and ORC-Associated (ORCA) are required for the initiation of DNA replication and regulate heterochromatin organization. Multiple subcomplexes of ORC and/or individual ORC subunits regulate different aspects of cell cycle progression and thus play pivotal roles in the maintenance of genomic stability. The long-term goal of my laboratory is to understand how ORC executes and coordinates various aspects of cell growth, proliferation and survival. The smallest subunit of ORC, Orc6, is required for DNA replication and also coordinates cytokinesis. The scientific premise of this proposal that human Orc6 is required for DNA replication progression and DNA Damage Response is based on strong preliminary data. We have exciting preliminary data demonstrating yet another novel role of Orc6, in DDR. We observe that the levels of Orc6 increase upon DNA damage and that Orc6 undergoes specific phosphorylation during oxidative DNA damage. Further, cells lacking Orc6 fail to activate ATR upon DNA damage. The objective of the present proposal is to answer fundamental questions on the roles of Orc6 in regulating S-phase and during DDR. Our hypothesis, based on preliminary data, is that Orc6 interacts with replication fork components and facilitates DNA replication and upon DNA damage the loss of this interaction inhibits replication fork progression. With our expertise and experience in cell biological and biochemical characterization of Orc6 and strong preliminary data, we are ideally positioned to pursue the following specific aims: 1) Determine the role of Orc6 in replication fork progression. 2) Determine the role of Orc6 in DNA damage response. 3) Determine the role of post-translational modifications on Orc6 in DDR and cell cycle progression. An important question in the field that has remained to be answered is why do ORC proteins remain bound to chromatin in postG1 cells? This proposal is significant because we address this question by studying how Orc6 regulates replication fork progression and it evaluates for the first time the role of preRC proteins in DNA damage response. This proposal is conceptually innovative because we will rigorously dissect novel regulatory mechanisms of Orc6 in DNA replication progression and DNA damage response. This proposal is technologically innovative because it employs state of the art cell biological techniques, including super-resolution imaging combined with biochemical and single molecule biophysical approaches. Understanding how Orc6 governs multiple pathways including DNA replication, mitosis and DDR is expected to uncover novel pathways that would be useful to prevent tumorigenesis and key to allow more effective therapeutic targeting to combat cancer.

基因组完整性的维持对细胞存活至关重要。精确复制我们的基因组， 对遗传毒性应激的适当细胞反应对于维持基因组稳定性是至关重要的。起源 识别复合物（ORC，由六个亚基组成）和ORC相关（ORCA）是 启动DNA复制和调节异染色质组织。的多重子复形 ORC和/或单个ORC亚基调节细胞周期进程的不同方面，从而发挥作用。 在维持基因组稳定性中起关键作用。我实验室的长期目标是 了解ORC如何执行和协调细胞生长，增殖和 生存ORC的最小亚基Orc 6是DNA复制所需的，也是协调 胞质分裂这一提议的科学前提是，人类Orc 6是DNA复制所必需的。 疾病进展和DNA损伤反应是基于强有力的初步数据。我们有令人兴奋 初步数据表明Orc 6在DDR中的另一种新作用。我们观察到， Orc 6在DNA损伤后增加，并且Orc 6在氧化过程中经历特异性磷酸化。 DNA损伤。此外，缺乏Orc 6的细胞在DNA损伤时不能激活ATR。的目的 目前的建议是回答关于Orc 6在调节S期中的作用的基本问题， 在DDR期间。基于初步数据，我们的假设是Orc 6与复制叉相互作用， 组分和促进DNA复制和DNA损伤后，这种相互作用的损失抑制 复制叉进展。凭借我们在细胞生物学和生物化学领域的专业知识和经验， Orc 6的特性和强大的初步数据，我们理想地定位于追求以下 具体目标：1）确定Orc 6在复制叉进展中的作用。2)确定Orc 6的角色 DNA损伤反应。3)确定Orc 6上的翻译后修饰在DDR中的作用， 细胞周期进程该领域一个尚待回答的重要问题是， ORC蛋白在G1后细胞中仍然与染色质结合？这一建议意义重大，因为我们 通过研究Orc 6如何调节复制叉进程来解决这个问题， 首次揭示了preRC蛋白在DNA损伤反应中的作用。该提案在概念上 创新，因为我们将严格剖析Orc 6在DNA复制中的新调控机制 DNA损伤反应。该提案具有技术创新性，因为它采用了 最先进的细胞生物学技术，包括超分辨率成像与生化 和单分子生物物理方法。了解Orc 6如何管理多个途径 包括DNA复制，有丝分裂和DDR，有望发现新的途径， 以防止肿瘤发生和关键允许更有效的治疗靶向对抗癌症。

项目成果

DNA Damage-Induced, S-Phase Specific Phosphorylation of Orc6 is Critical for the Maintenance of Genome Stability

• DOI: 10.1080/10985549.2023.2196204
• 发表时间: 2023-04
• 期刊: Molecular and Cellular Biology
• 影响因子: 5.3
• 作者: Yo-Chuen Lin;Dazhen Liu;Arindam Chakraborty;V. Macias;Eileen Brister;Jay Sonalkar;Linyuan Shen;Jaba Mitra;T. Ha;A. Kajdacsy-Balla;K. Prasanth;S. Prasanth

LncRNA-mediated regulation of SOX9 expression in basal subtype breast cancer cells

• DOI: 10.1261/rna.073254.119
• 发表时间: 2020-02-01
• 期刊: RNA
• 影响因子: 4.5
• 作者: Tariq, Aamira;Hao, Qinyu;Prasanth, Kannanganattu V.
• 通讯作者: Prasanth, Kannanganattu V.

MIR100 host gene-encoded lncRNAs regulate cell cycle by modulating the interaction between HuR and its target mRNAs.

• DOI: 10.1093/nar/gky696
• 发表时间: 2018-11-02
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Sun Q;Tripathi V;Yoon JH;Singh DK;Hao Q;Min KW;Davila S;Zealy RW;Li XL;Polycarpou-Schwarz M;Lehrmann E;Zhang Y;Becker KG;Freier SM;Zhu Y;Diederichs S;Prasanth SG;Lal A;Gorospe M;Prasanth KV
• 通讯作者: Prasanth KV

The E3 ligase RFWD3 stabilizes ORC in a p53-dependent manner

E3 连接酶 RFWD3 以 p53 依赖性方式稳定 ORC

• DOI: 10.1080/15384101.2020.1829823
• 发表时间: 2020
• 期刊: Cell Cycle
• 影响因子: 4.3
• 作者: Hsu, Rosaline Y.C.;Giri, Sumanprava;Wang, Yating;Lin, Yo-Chuen;Liu, Dazhen;Wopat, Susan;Chakraborty, Arindam;Prasanth, Kannanganattu V.;Prasanth, Supriya G.
• 通讯作者: Prasanth, Supriya G.

A BEN-domain protein and polycomb complex work coordinately to regulate transcription

• DOI: 10.1080/21541264.2022.2105128
• 发表时间: 2022-07-29
• 期刊: TRANSCRIPTION-AUSTIN
• 影响因子: 3.6
• 作者: Kurniawan, Fredy;Prasanth, Supriya G.
• 通讯作者: Prasanth, Supriya G.