Regulation of Replication Origin Usage in Saccharomyces cerevisiae
酿酒酵母复制起点使用的调控
基本信息
- 批准号:7391551
- 负责人:
- 金额:$ 28.07万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2006
- 资助国家:美国
- 起止时间:2006-04-01 至 2010-03-31
- 项目状态:已结题
- 来源:
- 关键词:Adverse effectsAffinityAtomic Force MicroscopyBindingBoxingCell AgingCell Cycle ProgressionCell ProliferationCell Proliferation RegulationCell divisionCellsChromatinChromatin StructureChromosome CondensationChromosomesComplexConditionDNA BindingDNA biosynthesisDNA chemical synthesisDNA-Directed DNA PolymeraseDefectDeoxyribonuclease IDeoxyribonucleasesDependenceElectron MicroscopyElectrophoretic Mobility Shift AssayElementsEtiologyFrequenciesGene ExpressionGenesGenomeGenomic InstabilityGenomicsHereditary DiseaseIn VitroLaboratoriesLarge T AntigenLengthLinkLocationMaintenanceMalignant NeoplasmsMapsMediatingModelingMolecularNucleosomesNumbersNutrientPatternPhasePlant RootsPlayPre-Replication ComplexProcessPropertyProteinsRangeRateRecruitment ActivityRegulationReplication InitiationReplication Initiation GeneReplication OriginResearch PersonnelRoleSaccharomyces cerevisiaeSimian virus 40SiteStressTestingTrisomyTumor Suppressor ProteinsYeastsautonomously replicating sequencebasecell growthchromatin immunoprecipitationcohesionhelicasein vivoinsightmutantorigin recognition complexprogramsresearch studytranscription factor
项目摘要
Coordination of DMA replication and gene expression is central to the regulation of cell proliferation.
A strategy for the coordination of these two processes is to engage the same regulators in both processes.
Classical examples of such dual functional regulators are the E. coll DnaA and the SV40 large T antigen,
which serve both the functions of regulators of replication initiation and gene expression. Mcm1 is a MADS
box transcription factor which regulates genes required for cell cycle progression and DMA replication. Its
activity is responsive to glycolytic flux, nutrient availability and environmental stresses. Mcm1 also binds
specific elements at replication origins to promote initiation of DMA replication. In this proposal, a direct
role for Mcm1 in the regulation of origin usage is investigated. The hypothesis that Mcm1 regulates origin
usage based on its occupancy under limiting conditions will be investigated at a genomic scale using three
different approaches: 1) to exhaustively isolate autonomously replicating sequences that are selectively
propagated, 2) to analyze the genome wide locations of Mcm1 at selected replication origins, 3) to identify
differentially activated early replicating chromosomal origins. Dependence of the recruitment/activation of
the pre-replication complex (pre-RC) on Mcm1 will be investigated by chromatin immunoprecipitation
experiments. Interactions between Mcm1 and components of the pre-RC will be analyzed by
electrophoretic mobility shift assay (EMSA) and Dnase! footprinting. Influence of Mcm1 on the local DMA
and chromatin structures will be visualized by atomic force microscopy, electron microscopy as well as
nucleosome mapping. Emerging examples of dual functional regulators that coordinate DMA replication and
gene expression during cell proliferation include E2F-RB and Myb-130. Modeling this strategy in yeast may
provide insights into the mechanistic actions of cell proliferation factors and tumor suppressors.
Mis-regulated DNA replication is known to have adverse effects ranging from uncontrolled cell
proliferation to cellular senescence. Uncoordinated DNA replication has also been linked to defects in
chromosome condensation, cohesion and fragmentation, all of which have dire consequences on genome
integrity. Therefore, understanding the regulation of DNA replication is central to the study of the etiology
of all genetic diseases rooted in genome instability including trisomy and cancer.
DMA复制和基因表达的协调是细胞增殖调节的核心。
协调这两个进程的一项战略是让同一监管机构参与这两个进程。
这种双功能调节剂的经典例子是E.科尔DNA A和SV 40大T抗原,
其起到复制起始和基因表达的调节剂的作用。M1是一个MADS
盒转录因子,调节细胞周期进程和DMA复制所需的基因。其
活性响应于糖酵解通量、营养可用性和环境压力。Mcm 1还结合
在复制起点的特定元件,以促进启动DMA复制。在这一建议中,一个直接
Mcm 1在原产地使用的调节作用进行了研究。Mcm 1调节起源的假说
基于其在限制条件下的占有率的使用将在基因组规模上使用三个
不同的方法:1)彻底分离选择性地被克隆的自主复制序列,
繁殖,2)分析Mcm 1在选定复制起点的全基因组位置,3)鉴定
差异激活的早期复制染色体起源。招募/激活的依赖性
将通过染色质免疫沉淀研究Mcm 1上复制前复合物(pre-RC
实验Mcm 1和预RC组分之间的相互作用将通过以下方法进行分析:
电泳迁移率变动分析(EMSA)和DNA酶!脚印Mcm 1对局部DMA的影响
染色质结构将通过原子力显微镜、电子显微镜以及
核小体作图新出现的协调DMA复制的双功能调节器的例子,
在细胞增殖期间表达的基因包括E2 F-RB和Myb-130。在酵母中模拟这种策略可能
提供深入了解细胞增殖因子和肿瘤抑制因子的机制作用。
已知错误调节的DNA复制具有不利影响,
增殖到细胞衰老。不协调的DNA复制也与基因缺陷有关。
染色体凝聚、凝聚和断裂,所有这些都对基因组产生可怕的后果
完整因此,了解DNA复制的调控是研究病因的核心
所有遗传疾病的根源在于基因组的不稳定性,包括三体和癌症。
项目成果
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{{ truncateString('BIK-KWOON TYE', 18)}}的其他基金
Regulation of Replication Origin Usage in Saccharomyces cerevisiae
酿酒酵母复制起点使用的调控
- 批准号:
7903070 - 财政年份:2009
- 资助金额:
$ 28.07万 - 项目类别:
Regulator of DNA Replication & Gene Expression in Yeast
DNA复制的调节者
- 批准号:
7088159 - 财政年份:2006
- 资助金额:
$ 28.07万 - 项目类别:
Regulation of Replication Origin Usage in Saccharomyces cerevisiae
酿酒酵母复制起点使用的调控
- 批准号:
7596349 - 财政年份:2006
- 资助金额:
$ 28.07万 - 项目类别:
Regulation of Replication Origin Usage in Saccharomyces cerevisiae
酿酒酵母复制起点使用的调控
- 批准号:
7197986 - 财政年份:2006
- 资助金额:
$ 28.07万 - 项目类别:
INTERACTIONS OF THE MCM PROTEINS AT REPLICATION ORIGINS
MCM 蛋白在复制起点的相互作用
- 批准号:
2177329 - 财政年份:1978
- 资助金额:
$ 28.07万 - 项目类别:
YEAST TRANSCRIPTION FACTOR INVOLVED IN DNA REPLICATION
参与 DNA 复制的酵母转录因子
- 批准号:
3284757 - 财政年份:1978
- 资助金额:
$ 28.07万 - 项目类别:
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