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The Molecular Logic of Lamellipodia Formation
The Molecular Logic of Lamellipodia Formation
상세정보
- 자료유형
- 학위논문 서양
- 최종처리일시
- 20260202103144
- ISBN
- 9798293851317
- DDC
- 574
- 저자명
- Wu, Muziyue.
- 서명/저자
- The Molecular Logic of Lamellipodia Formation
- 발행사항
- [Sl] : University of California, San Francisco, 2025
- 발행사항
- Ann Arbor : ProQuest Dissertations & Theses, 2025
- 형태사항
- 122 p
- 주기사항
- Source: Dissertations Abstracts International, Volume: 87-03, Section: B.
- 주기사항
- Advisor: Weiner, Orion.
- 학위논문주기
- Thesis (Ph.D.)--University of California, San Francisco, 2025.
- 초록/해제
- 요약Cells manipulate the shapes of their plasma membrane to execute different cellular functions, ranging from endocytosis, developmental morphogenesis to the protrusions that drive cell migration. All those processes are dependent on the rearrangement of the actin cytoskeleton, whose patterns of organization are controlled by proteins known as nucleation-promoting factors (NPFs). Cells use different NPFs to orchestrate dramatically different actin networks. For example, N-WASP controls the finger-like actin networks that comprise invadopodia, filopodia, and sites of endocytosis. In contrast, the WAVE regulatory complex organizes sheet-like protrusions known as lamellipodia that are ancient highly conserved engines of cell motility. The multivalent interactions that generate N-WASP organization are relatively well-understood and have been reconstituted with purified proteins in vitro. But how the WAVE complex oligomerizes into lines is not understood and has not proven amenable to biochemical reconstitution. This thesis project seeks to probe the rules of NPF organization that underlie lamellipodia formation and cell morphogenesis.In Chapter 2, we leveraged in vivo biochemical approaches to investigate how the native WAVE complex instructs the formation of sheet-like lamellipodia. We show that the WAVE complex is a core constituent of a linear multilayered protein array at the plasma membrane, expected for an NPF that builds sheet-like actin-based protrusions. Negative membrane curvature is both necessary and sufficient for WAVE complex linear membrane association in the presence of upstream activators(Rac, Arf1/6, PIP3) and the PRD domains of both WAVE2 and Abi2, providing a potential mechanistic basis for templating of lamellipodia and their emergent behaviors, including barrier avoidance. Through computational modeling we demonstrate that the WAVE complex's linear organization and preference for negative curvature both play important roles in robust lamellipodia formation. Our data reveal key features of mesoscale WAVE complex patterning and highlight an integral relation between NPF self-organization and cell morphogenesis.In Chapter 3, we leveraged synthetic biology to systematically probe how NPF organization leads to the emergent behaviors of cell morphogenesis and movement. We engineered synthetic NPFs that could mimic the organization pattern and function of the native NPFs. Those synthetic NPFs were designed to contain three modular parts: a protein scaffold that mimics the organization pattern of the native NPFs, a membrane binding motif to target to cell membrane, and an actin polymerization domain to build actin networks and initiate membrane morphogenesis. We tested the expression of a variety of de novo designed or native protein scaffolds in living cells, including the SPARK-ON droplets, microtubules, iPAK fibers, coiled coil sheets, and pseudobars, and observed desired localization and organization for some of those candidates. From there, we functionalized those protein scaffolds with actin polymerization domain, as well as incorporating optogenetic modules to achieve spatiotemporal control of actin polymerization and membrane morphogenesis. This synthetic approach enables us to test a much wider range of possible parameter spaces of NPF organization to probe how these biophysical properties link to function.In Chapter 4, I described my pilot work exploring and developing a variety of techniques and tools (nanotopography, DNA origami, polarized microscopy, and proximity labeling) that could be used to study the biophysical properties of native WAVE complex assembly as well as for validating and testing synthetic NPFs in the future.
- 일반주제명
- Cellular biology
- 일반주제명
- Molecular biology
- 일반주제명
- Biochemistry
- 일반주제명
- Biophysics
- 키워드
- Cell motility
- 키워드
- Cytoskeleton
- 키워드
- Plasma membrane
- 기타저자
- University of California, San Francisco Biophysics
- 기본자료저록
- Dissertations Abstracts International. 87-03B.
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
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■00520260202103144
■006m o d
■007cr#unu||||||||
■020 ▼a9798293851317
■035 ▼a(MiAaPQ)AAI31995159
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a574
■1001 ▼aWu, Muziyue.▼0(orcid)0000-0003-3096-8269
■24510▼aThe Molecular Logic of Lamellipodia Formation
■260 ▼a[Sl]▼bUniversity of California, San Francisco▼c2025
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2025
■300 ▼a122 p
■500 ▼aSource: Dissertations Abstracts International, Volume: 87-03, Section: B.
■500 ▼aAdvisor: Weiner, Orion.
■5021 ▼aThesis (Ph.D.)--University of California, San Francisco, 2025.
■520 ▼aCells manipulate the shapes of their plasma membrane to execute different cellular functions, ranging from endocytosis, developmental morphogenesis to the protrusions that drive cell migration. All those processes are dependent on the rearrangement of the actin cytoskeleton, whose patterns of organization are controlled by proteins known as nucleation-promoting factors (NPFs). Cells use different NPFs to orchestrate dramatically different actin networks. For example, N-WASP controls the finger-like actin networks that comprise invadopodia, filopodia, and sites of endocytosis. In contrast, the WAVE regulatory complex organizes sheet-like protrusions known as lamellipodia that are ancient highly conserved engines of cell motility. The multivalent interactions that generate N-WASP organization are relatively well-understood and have been reconstituted with purified proteins in vitro. But how the WAVE complex oligomerizes into lines is not understood and has not proven amenable to biochemical reconstitution. This thesis project seeks to probe the rules of NPF organization that underlie lamellipodia formation and cell morphogenesis.In Chapter 2, we leveraged in vivo biochemical approaches to investigate how the native WAVE complex instructs the formation of sheet-like lamellipodia. We show that the WAVE complex is a core constituent of a linear multilayered protein array at the plasma membrane, expected for an NPF that builds sheet-like actin-based protrusions. Negative membrane curvature is both necessary and sufficient for WAVE complex linear membrane association in the presence of upstream activators(Rac, Arf1/6, PIP3) and the PRD domains of both WAVE2 and Abi2, providing a potential mechanistic basis for templating of lamellipodia and their emergent behaviors, including barrier avoidance. Through computational modeling we demonstrate that the WAVE complex's linear organization and preference for negative curvature both play important roles in robust lamellipodia formation. Our data reveal key features of mesoscale WAVE complex patterning and highlight an integral relation between NPF self-organization and cell morphogenesis.In Chapter 3, we leveraged synthetic biology to systematically probe how NPF organization leads to the emergent behaviors of cell morphogenesis and movement. We engineered synthetic NPFs that could mimic the organization pattern and function of the native NPFs. Those synthetic NPFs were designed to contain three modular parts: a protein scaffold that mimics the organization pattern of the native NPFs, a membrane binding motif to target to cell membrane, and an actin polymerization domain to build actin networks and initiate membrane morphogenesis. We tested the expression of a variety of de novo designed or native protein scaffolds in living cells, including the SPARK-ON droplets, microtubules, iPAK fibers, coiled coil sheets, and pseudobars, and observed desired localization and organization for some of those candidates. From there, we functionalized those protein scaffolds with actin polymerization domain, as well as incorporating optogenetic modules to achieve spatiotemporal control of actin polymerization and membrane morphogenesis. This synthetic approach enables us to test a much wider range of possible parameter spaces of NPF organization to probe how these biophysical properties link to function.In Chapter 4, I described my pilot work exploring and developing a variety of techniques and tools (nanotopography, DNA origami, polarized microscopy, and proximity labeling) that could be used to study the biophysical properties of native WAVE complex assembly as well as for validating and testing synthetic NPFs in the future.
■590 ▼aSchool code: 0034.
■650 4▼aCellular biology
■650 4▼aMolecular biology
■650 4▼aBiochemistry
■650 4▼aBiophysics
■653 ▼aCell motility
■653 ▼aCytoskeleton
■653 ▼aMembrane morphogenesis
■653 ▼aSynthetic biology
■653 ▼aPlasma membrane
■690 ▼a0379
■690 ▼a0786
■690 ▼a0487
■690 ▼a0307
■71020▼aUniversity of California, San Francisco▼bBiophysics.
■7730 ▼tDissertations Abstracts International▼g87-03B.
■790 ▼a0034
■791 ▼aPh.D.
■792 ▼a2025
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17357184▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
