Conservation of chromatin recruitment mechanisms in metazoan DNA replication licensing factors

后生动物 DNA 复制许可因子中染色质招募机制的保守

• 批准号: 2308642
• 负责人: Matthew Parker
• 金额: $ 73.5万
• 依托单位: University of Texas Southwestern Medical Center
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2308642&HistoricalAwards=false
• 关键词: Conservation; chromatin; recruitment; mechanisms; metazoan

Life requires the faithful transmission of genetic information from parents to offspring through replication of genomic DNA. All animals must replicate their DNA, yet the proteins required for replication differ substantially between animals. This project will investigate similarities and differences in DNA replication across the animal kingdom by studying the process in evolutionarily ancient and more modern animals. The studies will extend from sea sponge to humans and should clarify how DNA replication is initiated and how evolution might result in differences in replication proteins while still maintaining their function. The project will also provide summer research opportunities to community college students who would not otherwise have the opportunity to train in an academic laboratory at an R1 institution. This broader impacts program is specifically aimed at increasing minority participation in science and encouraging student transition to four-year STEM degree and Ph.D. programs. The first step in DNA replication is loading of the replicative helicase, Mcm2-7 onto chromatin. This licensing process requires three proteins: Orc1-6, Cdc6, and Cdt1. Fly Orc1 possesses an intrinsically disordered region (IDR) that is necessary and sufficient for chromatin recruitment in vivo and mediates DNA-dependent phase separation in vitro. These activities are negatively regulated by phosphorylation. Interestingly, while all metazoan Orc1 orthologs possess IDRs, these regions are highly variable in length and lack linear sequence similarity, raising questions about functional conservation. Orc1 IDR orthologs do share similar amino acid composition, leading to the hypothesis that sequence composition may define the functions of this IDR class. This hypothesis will be tested by studies that investigate in vitro DNA binding and in vivo chromatin recruitment activity of different Orc1 IDR orthologs. The outcomes are expected to advance understanding of the conservation of replication licensing mechanisms and will also broadly inform the study of intrinsically disordered proteins by developing the concept of compositional homology, which may be applicable to many IDR classes.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

生活需要通过复制基因组DNA来忠实地将遗传信息从父母传播到后代。所有动物都必须复制其DNA，但是复制所需的蛋白质在动物之间有很大不同。该项目将通过研究进化上古老和更现代的动物的过程来研究整个动物界的DNA复制的相似性和差异。这些研究将从海绵扩展到人类，并应阐明如何启动DNA复制，以及进化如何导致复制蛋白的差异，同时仍保持其功能。该项目还将为社区大学生提供夏季的研究机会，他们否则将没有机会在R1机构的一家学术实验室进行培训。该更广泛的影响计划专门旨在增加对科学的少数参与，并鼓励学生过渡到四年的STEM学位和博士学位。程序。 DNA复制中的第一步是将复制性解旋酶（MCM2-7）加载到染色质上。此许可过程需要三种蛋白质：ORC1-6，CDC6和CDT1。 Fly ORC1具有本质上无序的区域（IDR），这对于体内的染色质募集是必要且足够的，并在体外介导了DNA依赖性相分离。这些活性受磷酸化负调节。有趣的是，尽管所有后生ORC1直系同源物都具有IDR，但这些区域的长度高度变化，缺乏线性序列相似性，从而提出了有关功能保护的问题。 ORC1 IDR直系同源物确实具有相似的氨基酸组成，从而假设序列组成可以定义该IDR类的功能。该假设将通过研究体外DNA结合和不同ORC1 IDR直系同源物的体内染色质募集活性的研究来检验。 The outcomes are expected to advance understanding of the conservation of replication licensing mechanisms and will also broadly inform the study of intrinsically disordered proteins by developing the concept of compositional homology, which may be applicable to many IDR classes.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

项目成果

Protein intrinsically disordered regions have a non-random, modular architecture.

• DOI: 10.1093/bioinformatics/btad732
• 发表时间: 2023-12-01
• 期刊: Bioinformatics (Oxford, England)

The origin recognition complex requires chromatin tethering by a hypervariable intrinsically disordered region that is functionally conserved from sponge to man

• DOI: 10.1093/nar/gkae122
• 发表时间: 2024-02-21
• 期刊: NUCLEIC ACIDS RESEARCH
• 影响因子: 14.9
• 作者: Adiji，Olubu A.;McConnell，Brendan S.;Parker，Matthew W.
• 通讯作者: Parker，Matthew W.