語系:
繁體中文
English
說明(常見問題)
回圖書館首頁
手機版館藏查詢
登入
回首頁
切換:
標籤
|
MARC模式
|
ISBD
FindBook
Google Book
Amazon
博客來
Structure-Based Studies of RNA Virus Entry and Assembly in the Native Context.
紀錄類型:
書目-電子資源 : Monograph/item
正題名/作者:
Structure-Based Studies of RNA Virus Entry and Assembly in the Native Context./
作者:
Chmielewski, David.
出版者:
Ann Arbor : ProQuest Dissertations & Theses, : 2022,
面頁冊數:
166 p.
附註:
Source: Dissertations Abstracts International, Volume: 84-01, Section: B.
Contained By:
Dissertations Abstracts International84-01B.
標題:
Viruses. -
電子資源:
http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=29176557
ISBN:
9798835549092
Structure-Based Studies of RNA Virus Entry and Assembly in the Native Context.
Chmielewski, David.
Structure-Based Studies of RNA Virus Entry and Assembly in the Native Context.
- Ann Arbor : ProQuest Dissertations & Theses, 2022 - 166 p.
Source: Dissertations Abstracts International, Volume: 84-01, Section: B.
Thesis (Ph.D.)--Stanford University, 2022.
This item must not be sold to any third party vendors.
Electron cryomicroscopy (cryo-EM) has rapidly become the method of choice for structural studies of macromolecular complexes in vitro, but often comes at the cost of losing valuable biological context related to structure/function in the native environment of the cell. Alphaviruses, a genus of positive-strand RNA enveloped viruses with significant human health burden, are generally variable in size and shape, and thus pose a challenge to traditional cryo-EM analysis. Further, structures important to alphavirus assembly/budding and replication processes are intricately linked to the context of infected cell membranes. While the released alphavirus virion architecture has been well characterized, the structural assembly mechanism of its two icosahedral protein layers remains undefined. In chapter 2 of this thesis, we analyze the structures of purified alphavirus particles using cryo-EM imaging and single particle analysis. We discovered a subpopulation of T=3 icosahedral particles, differing from the T=4 structure of all previous alphavirus structures. We then determined near-atomic resolution structures of Mayaro and Chikungunya virus, two circulating alphaviruses in the Americas, by addressing deviations from icosahedral symmetry within each particle. In chapter 3, we studied the assembly mechanism of alphavirus particles in situ using electron cryotomography (cryo-ET) of virus-infected cells. From snapshots of virus assembly/budding events, we determined 3D structures of assembly intermediates that revealed the mechanistic roles of glycoprotein spikes and nucleocapsids in forming two-layered icosahedral particles. Further, data of CHIKV-infected cells treated with budding-inhibiting antibodies shows that spacing spikes apart to prevent their lateral interactions prevents the plasma membrane bending around NC cores, thus blocking virus budding. These findings provide the molecular mechanisms of icosahedral enveloped virus assembly/budding and budding-blocking antibodies. Finally, we present an initial in situ structure of the functional CHIKV replication complex in virus-infected cells, with a preliminary model of viral non-structural protein organization in the cell. We suggest further analysis of this replication system will reveal the functional stages of positive-strand RNA processing in the cell.Lastly, in chapter 4, we switched focus to study the near-native structures of coronavirus spike (S) trimers directly on the surface of human coronavirus (HuCoV) virions in light of the Covid-19 pandemic. S trimers protruding from the viral envelope mediate coronavirus entry and are the main target for vaccine development. We first determined the 3.4A resolution structure of the alphacoronavirus HuCoV-NL63 prefusion S crown directly on virions without the need for recombinant, soluble protein with stabilizing mutations. Using further cryo-ET and subvolume analysis, we determined structural conformations of full-length S trimers, revealing incredible orientational flexibility mediated by a novel glycan hinge in the membrane-anchored stalk domain. Our results suggest a straightforward path for future studies of coronaviruses with external reagents such as receptors, drugs and antibodies, all in the context of pathogenicity relevant to human health.
ISBN: 9798835549092Subjects--Topical Terms:
571474
Viruses.
Structure-Based Studies of RNA Virus Entry and Assembly in the Native Context.
LDR
:04396nmm a2200337 4500
001
2352099
005
20221111121017.5
008
241004s2022 ||||||||||||||||| ||eng d
020
$a
9798835549092
035
$a
(MiAaPQ)AAI29176557
035
$a
(MiAaPQ)STANFORDjp217tq9449
035
$a
AAI29176557
040
$a
MiAaPQ
$c
MiAaPQ
100
1
$a
Chmielewski, David.
$3
3691719
245
1 0
$a
Structure-Based Studies of RNA Virus Entry and Assembly in the Native Context.
260
1
$a
Ann Arbor :
$b
ProQuest Dissertations & Theses,
$c
2022
300
$a
166 p.
500
$a
Source: Dissertations Abstracts International, Volume: 84-01, Section: B.
500
$a
Advisor: Chiu, Wah;Arvin, Ann;Carette, Jan;Sarnow, Peter.
502
$a
Thesis (Ph.D.)--Stanford University, 2022.
506
$a
This item must not be sold to any third party vendors.
520
$a
Electron cryomicroscopy (cryo-EM) has rapidly become the method of choice for structural studies of macromolecular complexes in vitro, but often comes at the cost of losing valuable biological context related to structure/function in the native environment of the cell. Alphaviruses, a genus of positive-strand RNA enveloped viruses with significant human health burden, are generally variable in size and shape, and thus pose a challenge to traditional cryo-EM analysis. Further, structures important to alphavirus assembly/budding and replication processes are intricately linked to the context of infected cell membranes. While the released alphavirus virion architecture has been well characterized, the structural assembly mechanism of its two icosahedral protein layers remains undefined. In chapter 2 of this thesis, we analyze the structures of purified alphavirus particles using cryo-EM imaging and single particle analysis. We discovered a subpopulation of T=3 icosahedral particles, differing from the T=4 structure of all previous alphavirus structures. We then determined near-atomic resolution structures of Mayaro and Chikungunya virus, two circulating alphaviruses in the Americas, by addressing deviations from icosahedral symmetry within each particle. In chapter 3, we studied the assembly mechanism of alphavirus particles in situ using electron cryotomography (cryo-ET) of virus-infected cells. From snapshots of virus assembly/budding events, we determined 3D structures of assembly intermediates that revealed the mechanistic roles of glycoprotein spikes and nucleocapsids in forming two-layered icosahedral particles. Further, data of CHIKV-infected cells treated with budding-inhibiting antibodies shows that spacing spikes apart to prevent their lateral interactions prevents the plasma membrane bending around NC cores, thus blocking virus budding. These findings provide the molecular mechanisms of icosahedral enveloped virus assembly/budding and budding-blocking antibodies. Finally, we present an initial in situ structure of the functional CHIKV replication complex in virus-infected cells, with a preliminary model of viral non-structural protein organization in the cell. We suggest further analysis of this replication system will reveal the functional stages of positive-strand RNA processing in the cell.Lastly, in chapter 4, we switched focus to study the near-native structures of coronavirus spike (S) trimers directly on the surface of human coronavirus (HuCoV) virions in light of the Covid-19 pandemic. S trimers protruding from the viral envelope mediate coronavirus entry and are the main target for vaccine development. We first determined the 3.4A resolution structure of the alphacoronavirus HuCoV-NL63 prefusion S crown directly on virions without the need for recombinant, soluble protein with stabilizing mutations. Using further cryo-ET and subvolume analysis, we determined structural conformations of full-length S trimers, revealing incredible orientational flexibility mediated by a novel glycan hinge in the membrane-anchored stalk domain. Our results suggest a straightforward path for future studies of coronaviruses with external reagents such as receptors, drugs and antibodies, all in the context of pathogenicity relevant to human health.
590
$a
School code: 0212.
650
4
$a
Viruses.
$3
571474
650
4
$a
Mosquitoes.
$3
1244706
650
4
$a
Coronaviruses.
$3
894828
650
4
$a
Mortality.
$3
533218
650
4
$a
Epidemics.
$3
588395
650
4
$a
Animal sciences.
$3
3174829
650
4
$a
Epidemiology.
$3
568544
650
4
$a
Medicine.
$3
641104
650
4
$a
Virology.
$3
642304
690
$a
0475
690
$a
0766
690
$a
0564
690
$a
0720
710
2
$a
Stanford University.
$3
754827
773
0
$t
Dissertations Abstracts International
$g
84-01B.
790
$a
0212
791
$a
Ph.D.
792
$a
2022
793
$a
English
856
4 0
$u
http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=29176557
筆 0 讀者評論
館藏地:
全部
電子資源
出版年:
卷號:
館藏
1 筆 • 頁數 1 •
1
條碼號
典藏地名稱
館藏流通類別
資料類型
索書號
使用類型
借閱狀態
預約狀態
備註欄
附件
W9474537
電子資源
11.線上閱覽_V
電子書
EB
一般使用(Normal)
在架
0
1 筆 • 頁數 1 •
1
多媒體
評論
新增評論
分享你的心得
Export
取書館
處理中
...
變更密碼
登入