Structure and Activity of DNA Replication Origins

DNA 复制起点的结构和活性

基本信息

批准号：
6765813
负责人：
DAVID KOWALSKI
金额：
$ 34.66万
依托单位：
ROSWELL PARK CANCER INSTITUTE CORP
依托单位国家：
美国
项目类别：
财政年份：
1983
资助国家：
美国
起止时间：
1983-03-01 至 2008-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6765813
关键词：
DNA footprinting DNA replication origin Saccharomyces cerevisiae chromosomes fungal genetics gene induction /repression gene mutation genetic regulatory element immunoprecipitation nucleic acid sequence nucleic acid structure regulatory gene site directed mutagenesis

项目摘要

DESCRIPTION (APPLICANT'S ABSTRACT): Activation and regulation of DNA replication are of fundamental importance to proper growth and differentiation of cells and development of organisms. Understanding how the molecular events that control DNA replication are altered by genetic mutations, environmental chemicals or therapeutic drugs is important in the prevention and treatment of diseases such as cancer. We will continue to use the yeast, Saccharomyces cerevisiae, to investigate how DNA replication is regulated in cell chromosomes. We have identified cis-acting DNA elements that are necessary and sufficient to specify and activate a chromosomal replication origin, 0RI305. One specific aim is to complete the characterization of a DNA unwinding element (DUE) present in 0RI305 and detected in the plasmid Autonomously Replicating Sequence, ARS3O5. We have discovered that silent replication origins near a transcriptionally-inactive chromosomal locus (HML) can be activated in the chromosome. One key component of origin silencing is the proximity of the nearest active origin in the chromosome. Our preliminary studies indicate that another important component is a chromosome position effect near HML. A specific aim is to identify DNA sequences and chromosome contexts responsible for the chromosome position effect that silences replication origins. Our preliminary studies indicate that, when activated, silent origins fire late in S phase. A specific aim is to identify the chromosome context that determines late replication timing at silent origins and to identify novel DNA elements that control late replication timing. Preliminary studies show that origin silencing is linked to the cellular response to DNA damage and that mutations in certain cell-cycle checkpoint genes activate silent origins. A specific aim is to examine the role of checkpoint genes and newly identified cis-acting mutations in controlling specific protein associations involved in origin silencing and late replication timing.

描述（申请人的摘要）：DNA的激活和调节复制对于适当的增长和差异至关重要细胞和生物的发育。了解分子事件控制DNA复制因遗传突变，环境而改变化学药品或治疗药物在预防和治疗中很重要癌症等疾病。我们将继续使用酵母，糖疗法酿酒酵母，研究如何在细胞中调节DNA复制染色体。我们已经确定了必要的顺式动作DNA元素，足以指定和激活染色体复制来源0RI305。一个具体的目的是完成DNA放松元素的表征（应得的）在0RI305中存在，并在质粒自主复制中检测到序列，ARS3O5。我们已经发现，沉默的复制起源附近转录活性染色体基因座（HML）可以在染色体。原点沉默的一个关键组成部分是染色体中最近的活性起源。我们的初步研究表明另一个重要组成部分是HML附近的染色体位置效应。一个具体目的是识别负责的DNA序列和染色体环境对于沉默复制起源的染色体位置效应。我们的初步研究表明，当激活时，无声的起源在后期发射 s阶段。一个具体的目的是确定决定的染色体上下文在沉默起源的晚期复制时机并识别新颖的DNA元素控制较晚的复制时间。初步研究表明起源沉默与对DNA损伤的细胞反应有关，该突变在某些细胞周期检查点基因激活无声来源。一个特定的目标是检查检查点基因的作用和新鉴定的顺式作用控制涉及的特定蛋白质关联的突变沉默和较晚的复制时间。

项目成果

期刊论文数量（26）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Replication fork stalling by bulky DNA damage: localization at active origins and checkpoint modulation.

DOI：
10.1093/nar/gkq1215
发表时间：
2011-04
期刊：
Nucleic acids research
影响因子：
14.9
作者：
Minca EC;Kowalski D
通讯作者：
Kowalski D

DNA replication forks pause at silent origins near the HML locus in budding yeast.

DNA 复制叉在芽殖酵母中 HML 基因座附近的沉默起点处暂停。

DOI：
10.1128/mcb.21.15.4938-4948.2001
发表时间：
2001
期刊：
Molecular and cellular biology
影响因子：
5.3
作者：
Wang,Y;Vujcic,M;Kowalski,D
通讯作者：
Kowalski,D

Mung bean nuclease cleavage of a dA + dT-rich sequence or an inverted repeat sequence in supercoiled PM2 DNA depends on ionic environment.

绿豆核酸酶对超螺旋 PM2 DNA 中富含 dA dT 的序列或反向重复序列的切割取决于离子环境。

DOI：
10.1093/nar/12.18.7087
发表时间：
1984
期刊：
Nucleic acids research
影响因子：
14.9
作者：
Sheflin,LG;Kowalski,D
通讯作者：
Kowalski,D

Thermal energy suppresses mutational defects in DNA unwinding at a yeast replication origin.

热能抑制酵母复制起点 DNA 解旋中的突变缺陷。

DOI：
10.1073/pnas.87.7.2486
发表时间：
1990
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Umek,RM;Kowalski,D
通讯作者：
Kowalski,D

Altered DNA conformations in the gene regulatory region of torsionally-stressed SV40 DNA.

扭转应力 SV40 DNA 基因调控区的 DNA 构象发生改变。