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Defining the Structural Mechanisms of DNA Replication Restart in E. coli Through Cryo-EM
Defining the Structural Mechanisms of DNA Replication Restart in E. coli Through Cryo-EM
상세정보
- 자료유형
- 학위논문 서양
- 최종처리일시
- 20260202105154
- ISBN
- 9798293839797
- DDC
- 574.191
- 저자명
- Ducos, Peter L.
- 서명/저자
- Defining the Structural Mechanisms of DNA Replication Restart in E. coli Through Cryo-EM
- 발행사항
- [Sl] : The University of Wisconsin - Madison, 2025
- 발행사항
- Ann Arbor : ProQuest Dissertations & Theses, 2025
- 형태사항
- 188 p
- 주기사항
- Source: Dissertations Abstracts International, Volume: 87-03, Section: B.
- 주기사항
- Advisor: Grant, Timothy.
- 학위논문주기
- Thesis (Ph.D.)--The University of Wisconsin - Madison, 2025.
- 초록/해제
- 요약Faithful duplication of the bacterial chromosome requires not only the initiation of DNA replication at defined origins but also the ability to restart replication at sites where replisomes dissociate prematurely. In Escherichia coli, this essential genome maintenance function is mediated by multiple replication restart pathways, including those directed by PriA and PriC. This thesis defines the structural mechanisms underpinning these distinct restart processes using single-particle cryogenic electron microscopy (cryo-EM).In the first study, we determined the structure of a PriA/PriB/replication fork complex, revealing how PriA engages both arms of the DNA fork and undergoes conformational remodeling to form a pore around lagging-strand DNA and expose a PriB-binding interface. These structural changes define a switch-like mechanism that links fork recognition to complex assembly, ensuring selective and robust restart initiation.The second study presents two structures of the complete PriA/PriB/DnaT/replication fork complex, providing the first full-length visualization of DnaT. These structures capture domain-specific oligomerization events on single-stranded DNA and reveal a mechanistic analogy between DnaT and the canonical initiator protein DnaA.Finally, a third study examines PriC-dependent restart, defining the structural basis for PriC-mediated recruitment of the DnaB/DnaC helicase-loader complex. These findings delineate an alternative, mechanistically distinct strategy for replisome reloading that operates independently of PriA.Collectively, these studies leverage cryo-EM's unique ability to resolve dynamic, compositionally heterogeneous complexes that have resisted traditional crystallographic approaches. By capturing multiple conformational states of key replication intermediates, this work establishes a comprehensive structural framework for replication restart in E. coli, revealing how divergent molecular strategies converge on a shared goal of maintaining replication continuity and genome integrity.
- 일반주제명
- Biophysics
- 일반주제명
- Biochemistry
- 일반주제명
- Genetics
- 키워드
- DNA replication
- 키워드
- DnaT
- 키워드
- PriA
- 기타저자
- The University of Wisconsin - Madison Biophysics
- 기본자료저록
- Dissertations Abstracts International. 87-03B.
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
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■00520260202105154
■006m o d
■007cr#unu||||||||
■020 ▼a9798293839797
■035 ▼a(MiAaPQ)AAI32242734
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a574.191
■1001 ▼aDucos, Peter L.
■24510▼aDefining the Structural Mechanisms of DNA Replication Restart in E. coli Through Cryo-EM
■260 ▼a[Sl]▼bThe University of Wisconsin - Madison▼c2025
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2025
■300 ▼a188 p
■500 ▼aSource: Dissertations Abstracts International, Volume: 87-03, Section: B.
■500 ▼aAdvisor: Grant, Timothy.
■5021 ▼aThesis (Ph.D.)--The University of Wisconsin - Madison, 2025.
■520 ▼aFaithful duplication of the bacterial chromosome requires not only the initiation of DNA replication at defined origins but also the ability to restart replication at sites where replisomes dissociate prematurely. In Escherichia coli, this essential genome maintenance function is mediated by multiple replication restart pathways, including those directed by PriA and PriC. This thesis defines the structural mechanisms underpinning these distinct restart processes using single-particle cryogenic electron microscopy (cryo-EM).In the first study, we determined the structure of a PriA/PriB/replication fork complex, revealing how PriA engages both arms of the DNA fork and undergoes conformational remodeling to form a pore around lagging-strand DNA and expose a PriB-binding interface. These structural changes define a switch-like mechanism that links fork recognition to complex assembly, ensuring selective and robust restart initiation.The second study presents two structures of the complete PriA/PriB/DnaT/replication fork complex, providing the first full-length visualization of DnaT. These structures capture domain-specific oligomerization events on single-stranded DNA and reveal a mechanistic analogy between DnaT and the canonical initiator protein DnaA.Finally, a third study examines PriC-dependent restart, defining the structural basis for PriC-mediated recruitment of the DnaB/DnaC helicase-loader complex. These findings delineate an alternative, mechanistically distinct strategy for replisome reloading that operates independently of PriA.Collectively, these studies leverage cryo-EM's unique ability to resolve dynamic, compositionally heterogeneous complexes that have resisted traditional crystallographic approaches. By capturing multiple conformational states of key replication intermediates, this work establishes a comprehensive structural framework for replication restart in E. coli, revealing how divergent molecular strategies converge on a shared goal of maintaining replication continuity and genome integrity.
■590 ▼aSchool code: 0262.
■650 4▼aBiophysics
■650 4▼aBiochemistry
■650 4▼aGenetics
■653 ▼aCryogenic electron microscopy
■653 ▼aDNA replication
■653 ▼aDnaT
■653 ▼aPriA
■653 ▼aBacterial chromosome
■690 ▼a0786
■690 ▼a0487
■690 ▼a0369
■71020▼aThe University of Wisconsin - Madison▼bBiophysics.
■7730 ▼tDissertations Abstracts International▼g87-03B.
■790 ▼a0262
■791 ▼aPh.D.
■792 ▼a2025
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17359661▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
