Mechanisms of DNA replication and maintenance in eukaryotes
Mechanisms of DNA replication and maintenance in eukaryotes
批准号:
10625860
负责人:
PETER M BURGERS
金额:
$69.74万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2016
资助国家:
美国
项目状态:
未结题
起止时间:
2016-06-01 至 2026-05-31
关键词:
ATR geneAddressBiochemicalBypassCell Cycle CheckpointCellsComplexCouplingCryoelectron MicroscopyDNADNA DamageDNA MaintenanceDNA biosynthesisDNA lesionDNA metabolismDNA replication forkDNA-Directed DNA PolymeraseDefectEnvironmentEukaryotaEukaryotic CellEventFunctional disorderGenetic studyHumanInvestigationLesionMalignant NeoplasmsMutagenesisOkazaki fragmentsOrthologous GenePathway interactionsPatientsProcessProtein KinaseRegulationReplication OriginRiskSaccharomyces cerevisiaeSystemTEL1 GeneYeastsbiophysical analysisfascinategenetic analysisgenetic informationinsightmodel organismresponsesensor
中文摘要
描述:本提案的重点是真核生物 DNA 复制机制的研究
细胞 (1) 以及复制功能障碍对自发性和损伤的影响-
诱导突变 (2),以及细胞周期检查点激活 (3)。我们的主要方法结合
对酿酒酵母进行生化和生物物理分析以及遗传分析,以获得
深入了解这三个广泛定义的途径及其相互关联性。我们的研究是
通过对在这些过程中发挥作用的复杂机械的结构研究进行增强。提议
DNA 复制研究基于滞后链机制定义方面的大量进展记录
DNA 复制和冈崎片段成熟。由于冈崎碎片代表了迄今为止最
所有细胞中都频繁出现 DNA 不连续性,因此有必要了解 DNA 的不同层次的调控
这个过程。冈崎片段成熟主要在明确的生化系统中进行研究,
与复制叉处发生的其他事件隔离。我们建议在以下范围内扩大我们的研究
完整的复制叉的背景,它已在酵母复制起点处组装。我们的
诱变研究将集中在主要参与者 DNA 聚合酶 z 和 Rev1 上。一方面,Rev1
促进 Pol z 绕过 DNA 损伤;另一方面,它通过抑制 Pol 来限制诱变的程度
病变狭窄环境外的 z 依赖性 DNA 合成。我们将揭开机制
这是 Rev1 双重调节功能的基础。我们检查点研究的主要重点是
两种传感器蛋白激酶 Mec1 和 Tel1,分别是人类 ATR 和 ATM 的直系同源物。
生化和遗传学研究将与冷冻电镜研究相结合,以了解基础如何
这些独特的蛋白激酶的活性被激活。此外,拥有高效的优势
DNA复制系统的可用将使我们能够开始解决复制停滞之间的耦合问题
以及下游反应途径。最后,根据 MIRA 原则,我们将追求其他
DNA 代谢中令人着迷的问题可能而且毫无疑问会在我们的研究过程中出现。
英文摘要
DESCRIPTION: This proposal is centered on studies of the mechanism of DNA replication in eukaryotic
cells (1) and on the consequences of replication dysfunction with regard to spontaneous and damage-
induced mutagenesis (2), and to cell cycle checkpoint activation (3). Our primary approaches combine
biochemical and biophysical analysis with genetic analysis in the yeast Saccharomyces cerevisiae, to gain
insight in each of these three broadly defined pathways and their interconnectivity. Our studies are
augmented with structural studies of the complex machineries that function in these processes. Proposed
DNA replication studies are based on a strong record of progress in defining mechanisms of lagging strand
DNA replication and Okazaki fragment maturation. Since Okazaki fragments represent by far the most
frequent DNA discontinuities in all cells, it is imperative to understand the different layers of regulation of
this process. Okazaki fragment maturation has primarily been studied in well-defined biochemical systems,
in isolation from other events that occur at the replication fork. We propose to expand our studies within the
context of a complete replication fork, which has been assembled at a yeast replication origin. Our
mutagenesis studies will center on the main actors, DNA polymerase z and Rev1. On the one hand, Rev1
promotes DNA lesion bypass by Pol z; on the other hand, it limits the extent of mutagenesis by inhibiting Pol
z-dependent DNA synthesis outside the narrow environment of the lesion. We will unravel the mechanism
that underlies this dual regulatory function of Rev1. The primary focus of our checkpoint studies is on the
two sensor protein kinases Mec1 and Tel1, the orthologs of human ATR and ATM, respectively.
Biochemical and genetic studies will be combined with cryo-EM studies to understand how the basal
activities of these unique protein kinases are activated. Furthermore, the advantage of having an efficient
DNA replication system available will allow us to begin addressing the coupling between replication arrest
and the downstream response pathways. Finally, in keeping with the MIRA principle, we will pursue other
fascinating questions in DNA metabolism that may, and undoubtedly will arise during our investigations.
期刊论文(21)
专著(0)
科研奖励(0)
会议论文
登录
查看更多内容
DOI:
10.1093/nar/gkab371
发表时间:
2021-06-04
期刊:
Nucleic acids research
影响因子:
14.9
作者:
[Kiktev DA, Dominska M, Zhang T, Dahl J, Stepchenkova EI, Mieczkowski P, Burgers PM, Lujan S, Burkholder A, Kunkel TA, Petes TD]
通讯作者:
Petes TD
DOI:
10.1002/bies.201700070
发表时间:
2017-08
期刊:
BioEssays : news and reviews in molecular, cellular and developmental biology
影响因子:
--
作者:
[Kunkel TA, Burgers PMJ]
通讯作者:
Burgers PMJ
DOI:
10.1093/nar/gkw1149
发表时间:
2017-02-17
期刊:
Nucleic acids research
影响因子:
14.9
作者:
[Kochenova OV, Bezalel-Buch R, Tran P, Makarova AV, Chabes A, Burgers PM, Shcherbakova PV]
通讯作者:
Shcherbakova PV
DOI:
10.1146/annurev-biochem-061516-044709
发表时间:
2017-06-20
期刊:
Annual review of biochemistry
影响因子:
16.6
作者:
[Burgers PMJ, Kunkel TA]
通讯作者:
Kunkel TA
DOI:
10.1074/jbc.m115.708263
发表时间:
2016-06-24
期刊:
The Journal of biological chemistry
影响因子:
--
作者:
[Sawicka M, Wanrooij PH, Darbari VC, Tannous E, Hailemariam S, Bose D, Makarova AV, Burgers PM, Zhang X]
通讯作者:
Zhang X
共 9 条
Mechanisms of DNA replication and maintenance in eukaryotes
-
批准号:9281879
-
项目类别:
-
资助金额:$67.52万
-
财政年份:2016
-
负责人:PETER M BURGERS
-
依托单位:
Mechanisms of DNA replication and maintenance in eukaryotes
-
批准号:10166037
-
项目类别:
-
资助金额:$69.74万
-
财政年份:2016
-
负责人:PETER M BURGERS
-
依托单位:
Mechanisms of DNA replication and maintenance in eukaryotes
-
批准号:10405648
-
项目类别:
-
资助金额:$69.74万
-
财政年份:2016
-
负责人:PETER M BURGERS
-
依托单位:
Mechanisms of DNA replication and maintenance in eukaryotes
-
批准号:9071509
-
项目类别:
-
资助金额:$48.15万
-
财政年份:2016
-
负责人:PETER M BURGERS
-
依托单位:
KINASE ACTIVATION IN THE DNA DAMAGE CHECKPOINTS
-
批准号:8669991
-
项目类别:
-
资助金额:$25.83万
-
财政年份:2009
-
负责人:PETER M BURGERS
-
依托单位:
KINASE ACTIVATION IN THE DNA DAMAGE CHECKPOINTS
-
批准号:9024552
-
项目类别:
-
资助金额:$25.83万
-
财政年份:2009
-
负责人:PETER M BURGERS
-
依托单位:
KINASE ACTIVATION IN THE DNA DAMAGE CHECKPOINTS
-
批准号:7997212
-
项目类别:
-
资助金额:$25.33万
-
财政年份:2009
-
负责人:PETER M BURGERS
-
依托单位:
KINASE ACTIVATION IN THE DNA DAMAGE CHECKPOINTS
-
批准号:8206589
-
项目类别:
-
资助金额:$25.33万
-
财政年份:2009
-
负责人:PETER M BURGERS
-
依托单位:
KINASE ACTIVATION IN THE DNA DAMAGE CHECKPOINTS
-
批准号:8503918
-
项目类别:
-
资助金额:$25.83万
-
财政年份:2009
-
负责人:PETER M BURGERS
-
依托单位:
KINASE ACTIVATION IN THE DNA DAMAGE CHECKPOINTS
-
批准号:7753226
-
项目类别:
-
资助金额:$25.58万
-
财政年份:2009
-
负责人:PETER M BURGERS
-
依托单位:
DNA REPLICATION IN YEAST
-
批准号:6665802
-
项目类别:
-
资助金额:$15.75万
-
财政年份:2002
-
负责人:PETER M BURGERS
-
依托单位:
DNA REPLICATION IN YEAST
-
批准号:6486682
-
项目类别:
-
资助金额:$15.75万
-
财政年份:2001
-
负责人:PETER M BURGERS
-
依托单位:
DNA REPLICATION IN YEAST
-
批准号:6336752
-
项目类别:
-
资助金额:$0.07万
-
财政年份:2000
-
负责人:PETER M BURGERS
-
依托单位:
STRUCTURE/FUNCTION OF YEAST DNA POLYMERASE DELTA
-
批准号:6138673
-
项目类别:
-
资助金额:$16.16万
-
财政年份:1999
-
负责人:PETER M BURGERS
-
依托单位:
STRUCTURE/FUNCTION OF YEAST DNA POLYMERASE DELTA
-
批准号:6490212
-
项目类别:
-
资助金额:$17.13万
-
财政年份:1999
-
负责人:PETER M BURGERS
-
依托单位:
STRUCTURE/FUNCTION OF YEAST DNA POLYMERASE DELTA
-
批准号:6343034
-
项目类别:
-
资助金额:$16.64万
-
财政年份:1999
-
负责人:PETER M BURGERS
-
依托单位:
STRUCTURE/FUNCTION OF YEAST DNA POLYMERASE DELTA
-
批准号:2729078
-
项目类别:
-
资助金额:$15.7万
-
财政年份:1999
-
负责人:PETER M BURGERS
-
依托单位:
ENZYMOLOGY OF REPLICATION OF YEAST CHROMOSOMAL DNA
-
批准号:6287290
-
项目类别:
-
资助金额:$34.72万
-
财政年份:1984
-
负责人:PETER M BURGERS
-
依托单位:
ENZYMOLOGY OF REPLICATION OF YEAST CHROMOSOMAL DNA
-
批准号:2749817
-
项目类别:
-
资助金额:$34.75万
-
财政年份:1984
-
负责人:PETER M BURGERS
-
依托单位:
ENZYMOLOGY OF REPLICATION OF YEAST CHROMOSOMAL DNA
-
批准号:6625031
-
项目类别:
-
资助金额:$34.65万
-
财政年份:1984
-
负责人:PETER M BURGERS
-
依托单位:
海外基金