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.

生命需要通过基因组DNA的复制将遗传信息从父母忠实地传递给后代。所有动物都必须复制它们的DNA，但复制所需的蛋白质在动物之间有很大差异。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动物王国中DNA复制的相似性和差异。这些研究将从海绵扩展到人类，并应澄清DNA复制是如何启动的，以及进化如何导致复制蛋白质的差异，同时仍保持其功能。该项目还将为社区学院的学生提供夏季研究机会，否则他们将没有机会在R1机构的学术实验室进行培训。这个更广泛的影响计划专门针对增加少数民族参与科学，并鼓励学生过渡到四年制STEM学位和博士学位。程序. DNA复制的第一步是将复制解旋酶Mcm 2 -7装载到染色质上。这个许可过程需要三种蛋白质：Orc 1 -6，Cdc 6和Cdt 1。Fly Orc 1具有一个内在无序区（IDR），该区域是体内染色质募集所必需和足够的，并在体外介导DNA依赖性相分离。这些活动受到磷酸化的负调控。有趣的是，虽然所有后生动物Orc 1直系同源物都具有IDR，但这些区域的长度高度可变，缺乏线性序列相似性，这引发了关于功能保守性的问题。Orc 1 IDR直系同源物确实具有相似的氨基酸组成，这导致了序列组成可能定义该IDR类的功能的假设。该假设将通过研究不同Orc 1 IDR直系同源物的体外DNA结合和体内染色质募集活性的研究进行检验。该成果预计将促进对复制许可机制的保护的理解，并通过发展可能适用于许多IDR类的组成同源性概念，广泛地为内在无序蛋白质的研究提供信息。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估来支持。

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.