Mechanistic studies of stalled DNA replication fork rescue
Mechanistic studies of stalled DNA replication fork rescue
批准号:
10387612
负责人:
Piero R Bianco
金额:
$6.02万
依托单位国家:
美国
项目类别:
财政年份:
2013
资助国家:
美国
项目状态:
已结题
起止时间:
2013-06-07 至 2022-04-30
关键词:
AffectAirAtomic Force MicroscopyAutomobile DrivingBinding ProteinsBiological ModelsBuffersCaringComplexCruciform DNADNADNA DamageDNA StructureDNA biosynthesisDNA lesionDNA replication forkDNA-Directed RNA PolymeraseDataDefectDevelopmentDiscriminationEnzymesEscherichia coliEukaryotaEventFailureFosteringGenetic RecombinationGenetic TranscriptionGenomeGenome StabilityGenomic InstabilityGoalsHealthLeadLesionMagnetismMalignant NeoplasmsMissionModelingMutationOrganismOutcomePathway interactionsPlayProcessProkaryotic CellsProteinsPublic HealthPublishingReactionResearchRoleSS DNA BPSpeedStructureTestingTimeTranslationsUnited States National Institutes of HealthWorkcombinatorialgenome integrityhelicasehomologous recombinationhuman diseaseimprovedin vivoinnovationinsightnovelparent grantpublic health relevancerepairedrestorationsingle moleculetemporal measurement
中文摘要
摘要:家长授权gm10056
英文摘要
ABSTRACT OF THE PARENT GRANT GM10056
There is a fundamental gap in understanding how stalled DNA replication forks are rescued. The continued existence of
this gap represents an important problem because, until it is filled, a complete and clear understanding of the mechanism of
stalled fork reactivation will be lacking. This understanding is crucial as defects in these repair mechanisms in higher
organisms lead to the accumulation of mutations leading to cancer, and the proposed studies are therefore directly relevant
to human disease. Consequently, the long-term goal is to understand the mechanism of stalled DNA replication fork
reactivation. The main objective of this proposal is to understand the interplay between the single-stranded DNA binding
protein (SSB) and key fork rescue enzymes on nucleoid templates and of the subsequent processing events leading to
restoration of a fork structure. To achieve this objective, this proposal is divided into three specific aims: 1), Determine the
mechanism(s) of fork regression; 2,) To determine how fork impediments affect fork regression; and 3), Ascertain the effects
of nucleoid-associated proteins on fork rescue enzymes. Under the first aim, magnetic tweezers and atomic force microscopy
(both in air and high-speed in buffer) will be used to determine how SSB loading and regression by RecG are affected by
PriA and to ascertain whether RecA and RuvAB are able to catalyze an efficient and unidirectional fork regression reaction.
When the proposed studies for Aim 1 are complete, a clear picture of the events at a nascent, stalled replication fork will be
provided. Under the second aim, the same two single DNA molecule approaches will be used to provide insight into the
effects of replisome impediments on stalled fork rescue, with high spatial and temporal resolution. At the conclusion of the
proposed studies for Aim 2, the effects of DNA lesions and protein-DNA complexes on fork rescue will be made clear and
it is anticipated that the mechanism(s) for displacing stalled RNA polymerase in the vicinity of forks will be obtained. Under
the final aim, magnetic tweezers to manipulate single molecules of DNA will be used to ascertain the effects of nucleoid-
associated proteins (NAPs) on fork rescue. When the proposed studies for Aim 3 are complete, it will be ascertained whether
NAPs catalyze regression on their own and if they assist or inhibit fork rescue enzymes. The proposed research is innovative
because of the combinatorial strategy taken. It is also innovative because of the exciting and novel single-molecule
approaches used, the focus on nucleoid templates, and an understanding to be gained of how the primary protein barrier(s)
causing replisome stalling are removed. Finally, the work is also innovative because of the care taken in elucidating how
recombination helicases function in the presence of SSB. The proposed research is significant because it will allow, for the
first time, the development of clear models of the mechanistic events occurring at a stalled fork embedded within nucleoid
templates and, it will provide the first real-time insight into the range of events that transpire to reactivate a stalled fork in
vivo.
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Helicase unwinding: active or merely perfect?
解旋酶解旋:主动还是仅仅完美?
DOI:
10.1016/j.jmb.2012.04.030
发表时间:
2012
期刊:
Journal of molecular biology
影响因子:
5.6
作者:
[Bianco,PieroR, Webb,MartinR]
通讯作者:
Webb,MartinR
DOI:
10.1039/c6nr06850b
发表时间:
2016-12-07
期刊:
Nanoscale
影响因子:
6.7
作者:
[Zhang Y, Hashemi M, Lv Z, Lyubchenko YL]
通讯作者:
Lyubchenko YL
DOI:
10.3389/fmolb.2022.784451
发表时间:
2022
期刊:
Frontiers in molecular biosciences
影响因子:
5
作者:
[Bianco PR]
通讯作者:
Bianco PR
Characterize the Interaction of the DNA Helicase PriA with the Stalled DNA Replication Fork Using Atomic Force Microscopy.
使用原子力显微镜表征 DNA 解旋酶 PriA 与停滞的 DNA 复制叉的相互作用。
DOI:
10.21769/bioprotoc.3940
发表时间:
2021
期刊:
Bio-protocol
影响因子:
0.8
作者:
[Wang,Yaqing, Sun,Zhiqiang, Bianco,PieroR, Lyubchenko,YuriL]
通讯作者:
Lyubchenko,YuriL
DOI:
10.1615/critreveukaryotgeneexpr.v26.i1.70
发表时间:
2016
期刊:
Critical reviews in eukaryotic gene expression
影响因子:
1.6
作者:
[Lyubchenko YL, Shlyakhtenko LS]
通讯作者:
Shlyakhtenko LS
共 27 条
Insight into the mechanism of action of the SSB interactome
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批准号:10340457
-
项目类别:
-
资助金额:$37.86万
-
财政年份:2022
-
负责人:Piero R Bianco
-
依托单位:
Insight into the mechanism of action of the SSB interactome
-
批准号:10610679
-
项目类别:
-
资助金额:$22.87万
-
财政年份:2022
-
负责人:Piero R Bianco
-
依托单位:
Mechanistic studies of stalled DNA replication fork rescue
-
批准号:8439481
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项目类别:
-
资助金额:$51.87万
-
财政年份:2013
-
负责人:Piero R Bianco
-
依托单位:
Mechanistic studies of stalled DNA replication fork rescue
-
批准号:9263530
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项目类别:
-
资助金额:$11.81万
-
财政年份:2013
-
负责人:Piero R Bianco
-
依托单位:
Mechanistic studies of stalled DNA replication fork rescue
-
批准号:10291961
-
项目类别:
-
资助金额:$45.14万
-
财政年份:2013
-
负责人:Piero R Bianco
-
依托单位:
Mechanistic studies of stalled DNA replication fork rescue
-
批准号:8812888
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项目类别:
-
资助金额:$44.0万
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财政年份:2013
-
负责人:Piero R Bianco
-
依托单位:
Mechanistic studies of stalled DNA replication fork rescue
-
批准号:8667479
-
项目类别:
-
资助金额:$43.93万
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财政年份:2013
-
负责人:Piero R Bianco
-
依托单位:
Single Molecule Studies of DNA Helicases
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批准号:6557516
-
项目类别:
-
资助金额:$40.96万
-
财政年份:2003
-
负责人:Piero R Bianco
-
依托单位:
Single Molecule Studies of DNA Helicases
-
批准号:6840389
-
项目类别:
-
资助金额:$26.97万
-
财政年份:2003
-
负责人:Piero R Bianco
-
依托单位:
Single Molecule Studies of DNA Helicases
-
批准号:7002229
-
项目类别:
-
资助金额:$27.12万
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财政年份:2003
-
负责人:Piero R Bianco
-
依托单位:
Single Molecule Studies of DNA Helicases
-
批准号:6692214
-
项目类别:
-
资助金额:$26.2万
-
财政年份:2003
-
负责人:Piero R Bianco
-
依托单位:
Single Molecule Studies of DNA Helicases
-
批准号:7163528
-
项目类别:
-
资助金额:$27.12万
-
财政年份:2003
-
负责人:Piero R Bianco
-
依托单位:
国内基金
海外基金
湍流和化学交互作用对H2-Air-H2O微混燃烧中NO生成的影响研究
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批准号:51976048
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项目类别:面上项目
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资助金额:61.0万元
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批准年份:2019
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负责人:邱朋华
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依托单位: