서브메뉴
검색
Identification and Characterization of Novel Eukaryotic Chaperones
Identification and Characterization of Novel Eukaryotic Chaperones
상세정보
- 자료유형
- 학위논문 서양
- 최종처리일시
- 20250211151425
- ISBN
- 9798382784229
- DDC
- 574
- 저자명
- Nelliat, Anjali.
- 서명/저자
- Identification and Characterization of Novel Eukaryotic Chaperones
- 발행사항
- [Sl] : Harvard University, 2024
- 발행사항
- Ann Arbor : ProQuest Dissertations & Theses, 2024
- 형태사항
- 146 p
- 주기사항
- Source: Dissertations Abstracts International, Volume: 85-12, Section: B.
- 주기사항
- Advisor: Denic, Vladimir.
- 학위논문주기
- Thesis (Ph.D.)--Harvard University, 2024.
- 초록/해제
- 요약Protein folding and assembly is aided by molecular chaperones that prevent aggregation of unfolded or partially folded intermediates and guide them to their native folded state. Chaperones engage clients through cycles of binding and release from aggregation-prone, non-native regions. ATP-dependent chaperones utilize ATP hydrolysis to drive chaperone conformational changes between low and high affinity states, while ATP-independent chaperones are regulated by diverse mechanisms.Zinc-finger protein 1 (Zpr1) is an essential ATP-independent chaperone dedicated to the biogenesis of eukaryotic translation elongation factor 1A (eEF1A), a highly abundant GTP-binding protein. How Zpr1-mediated folding is regulated to ensure rapid Zpr1 recycling remains an unanswered question. We identified the highly conserved altered inheritance of mitochondria 29 (Aim29) as an eEF1A biogenesis factor. Structural modeling using AlphaFold-Multimer suggested that Aim29 senses the GTP-bound conformation of eEF1A folding intermediates bound to Zpr1. We validated this prediction using yeast genetics, cell biological and biochemical reconstitution approaches, and uncovered that Aim29 sensing of GTP-bound eEF1A coupled to a GTP hydrolysis event facilitates eEF1A exit from the folding cycle and allows for Zpr1 recycling. Our work reveals that a bespoke ATP-independent chaperone system has mechanistic similarity to ATPase chaperones, but unexpectedly relies on client GTP hydrolysis to regulate the chaperone-client interaction.Next, we attempted to use AlphaFold-Multimer to identify additional chaperones or biogenesis factors that haven't yet been uncovered. We optimized an Alphafold-based pipeline for screening a protein of interest against the entire yeast proteome to identify high-confidence interactors. The success of this screening approach is highlighted by two examples. First, we identified the previously uncharacterized but conserved eukaryotic protein Ypl225w as an eEF1A chaperone candidate and subsequent work by another graduate student in the lab revealed that Ypl225w was a ribosome-associating chaperone that mediates GTP-driven vectorial folding of nascent eEF1A. We also applied this pipeline to an essential eukaryotic GTPase of unknown function, Drosophila melanogaster Misato-Like protein (Dml1). The top interactors for Dml1 were subunits of the chaperonin-containing T-complex (CCT), which we validated experimentally. Acute depletion of Dml1 lead to a decrease in levels of assembled CCT and accumulation of monomers. This observation, along with structural modeling and other preliminary results suggest that Dml1 could be an assembly chaperone for CCT.
- 일반주제명
- Biology
- 일반주제명
- Bioinformatics
- 일반주제명
- Biochemistry
- 키워드
- Chaperones
- 키워드
- Protein folding
- 키워드
- Proteostasis
- 기타저자
- Harvard University Systems Biology
- 기본자료저록
- Dissertations Abstracts International. 85-12B.
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
008250123s2024 us c eng d■001000017161644
■00520250211151425
■006m o d
■007cr#unu||||||||
■020 ▼a9798382784229
■035 ▼a(MiAaPQ)AAI31294615
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a574
■1001 ▼aNelliat, Anjali.▼0(orcid)0000-0003-1344-5423
■24510▼aIdentification and Characterization of Novel Eukaryotic Chaperones
■260 ▼a[Sl]▼bHarvard University▼c2024
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2024
■300 ▼a146 p
■500 ▼aSource: Dissertations Abstracts International, Volume: 85-12, Section: B.
■500 ▼aAdvisor: Denic, Vladimir.
■5021 ▼aThesis (Ph.D.)--Harvard University, 2024.
■520 ▼aProtein folding and assembly is aided by molecular chaperones that prevent aggregation of unfolded or partially folded intermediates and guide them to their native folded state. Chaperones engage clients through cycles of binding and release from aggregation-prone, non-native regions. ATP-dependent chaperones utilize ATP hydrolysis to drive chaperone conformational changes between low and high affinity states, while ATP-independent chaperones are regulated by diverse mechanisms.Zinc-finger protein 1 (Zpr1) is an essential ATP-independent chaperone dedicated to the biogenesis of eukaryotic translation elongation factor 1A (eEF1A), a highly abundant GTP-binding protein. How Zpr1-mediated folding is regulated to ensure rapid Zpr1 recycling remains an unanswered question. We identified the highly conserved altered inheritance of mitochondria 29 (Aim29) as an eEF1A biogenesis factor. Structural modeling using AlphaFold-Multimer suggested that Aim29 senses the GTP-bound conformation of eEF1A folding intermediates bound to Zpr1. We validated this prediction using yeast genetics, cell biological and biochemical reconstitution approaches, and uncovered that Aim29 sensing of GTP-bound eEF1A coupled to a GTP hydrolysis event facilitates eEF1A exit from the folding cycle and allows for Zpr1 recycling. Our work reveals that a bespoke ATP-independent chaperone system has mechanistic similarity to ATPase chaperones, but unexpectedly relies on client GTP hydrolysis to regulate the chaperone-client interaction.Next, we attempted to use AlphaFold-Multimer to identify additional chaperones or biogenesis factors that haven't yet been uncovered. We optimized an Alphafold-based pipeline for screening a protein of interest against the entire yeast proteome to identify high-confidence interactors. The success of this screening approach is highlighted by two examples. First, we identified the previously uncharacterized but conserved eukaryotic protein Ypl225w as an eEF1A chaperone candidate and subsequent work by another graduate student in the lab revealed that Ypl225w was a ribosome-associating chaperone that mediates GTP-driven vectorial folding of nascent eEF1A. We also applied this pipeline to an essential eukaryotic GTPase of unknown function, Drosophila melanogaster Misato-Like protein (Dml1). The top interactors for Dml1 were subunits of the chaperonin-containing T-complex (CCT), which we validated experimentally. Acute depletion of Dml1 lead to a decrease in levels of assembled CCT and accumulation of monomers. This observation, along with structural modeling and other preliminary results suggest that Dml1 could be an assembly chaperone for CCT.
■590 ▼aSchool code: 0084.
■650 4▼aBiology
■650 4▼aBioinformatics
■650 4▼aBiochemistry
■653 ▼aChaperones
■653 ▼aProtein folding
■653 ▼aProteostasis
■653 ▼aTranslation elongation factors
■653 ▼aEukaryotic translation
■690 ▼a0306
■690 ▼a0715
■690 ▼a0487
■71020▼aHarvard University▼bSystems Biology.
■7730 ▼tDissertations Abstracts International▼g85-12B.
■790 ▼a0084
■791 ▼aPh.D.
■792 ▼a2024
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17161644▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
