Mechanisms of heterochromatin replication

异染色质复制机制

基本信息

批准号：
10078281
负责人：
Marcus Smolka
金额：
$ 30.11万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-01-01 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10078281
关键词：
Acetyltransferase Binding Biochemical Biological Cell Proliferation Cell physiology Cells Centromere Chromatin Complex Couples Coupling DNA Damage DNA biosynthesis DNA lesion Genome Genomic Instability Genomic approach Genomics HAT1 gene Heterochromatin Histone Acetylation Histones Human Imaging Techniques Knowledge Link Location Maintenance Malignant Neoplasms Malignant neoplasm of ovary Measures Mediator of activation protein Methylation Methyltransferase Mitosis Molecular Monitor Multiple Myeloma Mutation Nature Proliferating Protein Family Proteomics Regulation Replication Initiation Role S Phase S-Adenosylmethionine Satellite DNA Site Somatic Cell System Techniques Testing Therapeutic Time cancer cell cancer type density experimental study genome integrity genomic locus histone acetyltransferase insight member new therapeutic target novel overexpression prevent telomere virtual

项目摘要

PROJECT SUMMARY Heterochromatic domains, such as centromeres, telomeres and other satellite DNA, pose a major challenge for DNA replication. Compacted chromatin is thought to inhibit replication initiation and obstruct progression of the replication machinery. Not surprisingly, recent analyses of various somatic and cancer cells have revealed that repressive chromatin is associated with regions of late replication and high mutation density. Little, however, is known about the molecular mechanisms that control and facilitate DNA replication at heterochromatic domains. Of importance, cancer cells may harness these mechanisms to facilitate heterochromatin replication and sustain their increased proliferative demands. For example, overexpression of KDM4A/JMJD2, a demethylase that removes the heterochromatic mark H3K9 tri-methylation, enables chromatin de-compaction and accelerated replication in ovarian cancers. Strikingly, KDM4A overexpression also resulted in copy gain of specific genomic loci often amplified in ovarian cancers and multiple myeloma, further strengthening the link between de-regulated heterochromatin replication and genomic instability. These observations highlight the need to elucidate the mechanisms required for proper heterochromatin replication so as to understand fundamental aspects of genome maintenance and the control of cell proliferation. This proposal is centered on METTL13 (Methyltransferase-like 13), a member of a poorly understood family of proteins containing putative SAM (S-adenosylmethionine)-binding domains. While METTL13 was found amplified and overexpressed in cancers, virtually nothing was previously known about its cellular functions. Preliminary results presented here provide the first insights on how METTL13 sustains cell proliferation, revealing crucial roles in DNA replication and chromatin dynamics. More specifically, we find that METTL13 is a novel key mediator of heterochromatin replication that is particularly important for replication of centromeres. To the best of our knowledge this is the first described regulator of human centromere replication timing. This proposal will combine cutting edge genomic and proteomic techniques with biochemical and cell biological approaches to dissect the action of METTL13 and establish its role in controlling chromatin dynamics, DNA replication and genome integrity. Generated results will reveal a fundamental mechanism of heterochromatin replication and replication timing control, and will establish novel drug targets for modulating chromatin dynamics and the proliferative capacity of cancer cells.

项目概要异染色质结构域，如着丝粒、端粒和其他卫星 DNA，对 DNA复制。压缩的染色质被认为可以抑制复制起始并阻碍复制的进展。复制机器。毫不奇怪，最近对各种体细胞和癌细胞的分析表明，抑制性染色质与复制晚期和高突变密度区域相关。然而，很少的是了解控制和促进异染色质域 DNA 复制的分子机制。重要的是，癌细胞可以利用这些机制来促进异染色质复制和维持其日益增长的增殖性需求。例如，KDM4A/JMJD2（一种去甲基酶）的过度表达去除异染色质标记 H3K9 三甲基化，使染色质解压缩并加速卵巢癌的复制。引人注目的是，KDM4A 过度表达还导致复制增益特定基因组位点通常在卵巢癌和多发性骨髓瘤中扩增，进一步加强了这种联系异染色质复制失调和基因组不稳定之间的关系。这些观察结果凸显了需要阐明正确异染色质复制所需的机制，以便了解基因组维护和细胞增殖控制的基本方面。该提案以 METTL13（类甲基转移酶 13）为中心，它是一个人们知之甚少的成员。包含假定的 SAM（S-腺苷甲硫氨酸）结合域的蛋白质家族。虽然 METTL13 是发现在癌症中扩增和过度表达，之前几乎对其细胞一无所知功能。这里提供的初步结果提供了关于 METTL13 如何维持细胞的初步见解增殖，揭示了 DNA 复制和染色质动力学中的关键作用。更具体地说，我们发现 METTL13 是异染色质复制的新型关键介质，对于异染色质复制尤其重要着丝粒。据我们所知，这是第一个被描述的人类着丝粒调节因子复制时间。该提案将结合最先进的基因组和蛋白质组技术生物化学和细胞生物学方法来剖析 METTL13 的作用并确定其在控制染色质动力学、DNA 复制和基因组完整性。生成的结果将显示异染色质复制和复制时间控制的基本机制，并将建立新的调节染色质动力学和癌细胞增殖能力的药物靶点。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A field guide to the proteomics of post-translational modifications in DNA repair.

DOI：
10.1002/pmic.202200064
发表时间：
2022-08
期刊：
Proteomics
影响因子：
3.4
作者：
通讯作者：

Maximized quantitative phosphoproteomics allows high confidence dissection of the DNA damage signaling network.

DOI：
10.1038/s41598-020-74939-4
发表时间：
2020-10-22
期刊：
Scientific reports
影响因子：
4.6
作者：
Faca VM;Sanford EJ;Tieu J;Comstock W;Gupta S;Marshall S;Yu H;Smolka MB
通讯作者：
Smolka MB

Fe-NTA Microcolumn Purification of Phosphopeptides from Immunoprecipitation (IP) Eluates for Mass Spectrometry Analysis.

Fe-NTA 微柱纯化免疫沉淀 (IP) 洗脱液中的磷酸肽，用于质谱分析。

DOI：
10.21769/bioprotoc.4113
发表时间：
2021
期刊：
Bio-protocol
影响因子：
0.8
作者：
Sanford,EthanJ;Smolka,MarcusB
通讯作者：
Smolka,MarcusB

Characterization of an anti-FLAG antibody binding protein in V. cholerae.

霍乱弧菌中抗 FLAG 抗体结合蛋白的表征。

DOI：
10.1016/j.bbrc.2020.05.169
发表时间：
2020
期刊：
Biochemical and biophysical research communications
影响因子：
3.1
作者：
Shin,Jung-Ho;Lanz,Michael;Smolka,MarcusB;Dörr,Tobias
通讯作者：
Dörr,Tobias

Checkpoint-mediated DNA polymerase ε exonuclease activity curbing counteracts resection-driven fork collapse.

DOI：
10.1016/j.molcel.2021.04.006
发表时间：
2021-07-01
期刊：
Molecular cell
影响因子：
16
作者：
Pellicanò G;Al Mamun M;Jurado-Santiago D;Villa-Hernández S;Yin X;Giannattasio M;Lanz MC;Smolka MB;Yeeles J;Shirahige K;García-Díaz M;Bermejo R
通讯作者：
Bermejo R