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Molecular mechanisms of attachment by the ventral disc in Giardia

贾第鞭毛虫腹盘附着的分子机制

基本信息

批准号：
10552607
负责人：
SCOTT C DAWSON
金额：
$ 52.14万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-03-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10552607
关键词：
3-Dimensional Acute Diarrhea Affect Animal Model Animals Architecture Biological Biological Assay Biological Testing Biophysics CRISPR interference Cells Chronic diarrhea Clustered Regularly Interspaced Short Palindromic Repeats Complex Cyst Cytoplasm Data Defect Development Dorsal Electron Microscopy Elements Flagella Foundations Genetic Gerbils Giardia Image In Vitro Infection Intestinal parasite Intestines Knock-out Lateral Lead Life Cycle Stages Light Link Mediating Microscopy Microtubule Stabilization Microtubules Modeling Molecular Molecular Conformation Movement Organelles Parasites Peristalsis Persons Phenotype Proteins Research Resistance Resolution Role Severities Side Small Intestines Structural defect Structure Suction Surface Testing Theoretical model Time Work Zoonoses arm bioluminescence imaging cell motility diarrheal disease evidence base flexibility gut colonization host colonization hydrodynamic model imaging study in vivo insight knock-down link protein mutant novel prevent protein complex seal targeted treatment tool

项目摘要

Giardia is a widespread zoonotic intestinal parasite that causes acute and chronic diarrheal disease in more than 280 million people each year. Motile trophozoites colonize and attach to the small intestine with the ventral disc, a complex microtubule organelle. Attachment is required for infection as it allows Giardia to resist peristalsis. Theoretical models of attachment must be grounded in accurate biological data. For 50 years, hydrodynamic suction has been extensively modeled as the leading mechanism for Giardia attachment, yet this model of attachment lacks empirical support. Our pioneering work on disc architecture and composition, combined with our development of CRISPR-mediated knockdowns and knockouts and bioluminescent imaging of infection dynamics in animals, enable us to genetically test the structural and/or functional roles of DAPs required for disc conformational dynamics in attachment. During early stages of attachment, we discovered that regions of the disc undergo specific conformational changes. These changes, along with the presence of a curved disc, likely create a “seal” that enables attachment to the surface and resistance to shear forces. We have also recently identified and localized 87 disc-associated proteins (DAPs) to the specific structural and functional regions of the disc involved in maintaining and modulating disc conformations. Here we use our new CRISPR-based genetic tools to create specific classes of DAP mutants associated with key regions of the ventral disc that are likely required for its domed structure, as well as DAP mutants in key regions associated with flexible movements. We evaluate structural and attachment defects in CRISPR-interference (CRISPRi) DAP knockdowns (10 total) using high-resolution live imaging, electron microscopy, and biophysical assays. In Aim 1, we interrogate the role of the conspicuous microribbon and crossbridge complexes of the disc in mediating disc curvature. In Aim 2, we define the structural and functional roles of overlap zone DAPs that may structurally link the upper and lower portions of the disc, enabling proper domed conformation. In Aim 3, we interrogate the molecular mechanisms of DAPs associated with disc margin and ventral groove movements that contribute to formation of the lateral crest seal needed to resist shear forces. Lastly, we evaluate aberrant infection dynamics of two disc structural mutants using in vivo bioluminescent imaging (BLI) in an animal model of infection. Therapies that target parasite attachment would limit host colonization and limit the dissemination of infectious cysts.

贾第鞭毛虫是一种广泛传播的人畜共患肠道寄生虫，可引起急性和慢性腹泻

项目成果

期刊论文数量（22）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Eight unique basal bodies in the multi-flagellated diplomonad Giardia lamblia.

DOI：
10.1186/s13630-016-0042-4
发表时间：
2016-01-01
期刊：
Cilia
影响因子：
0
作者：
McInally, Shane G;Dawson, Scott C
通讯作者：
Dawson, Scott C

Mad2, Bub3, and Mps1 regulate chromosome segregation and mitotic synchrony in Giardia intestinalis, a binucleate protist lacking an anaphase-promoting complex.

DOI：
10.1091/mbc.e14-05-0975
发表时间：
2014-09-15
期刊：
Molecular biology of the cell
影响因子：
3.3
作者：
Vicente JJ;Cande WZ
通讯作者：
Cande WZ

'Disc-o-Fever': Getting Down with Giardia's Groovy Microtubule Organelle.

“Disc-o-Fever”：了解贾第虫的 Groovy 微管细胞器。

DOI：
10.1016/j.tcb.2017.10.007
发表时间：
2018
期刊：
Trends in cell biology
影响因子：
19
作者：
Nosala,Christopher;Hagen,KariD;Dawson,ScottC
通讯作者：
Dawson,ScottC

Recent advances in functional research in Giardia intestinalis.

DOI：
10.1016/bs.apar.2019.12.002
发表时间：
2020
期刊：
Advances in parasitology
影响因子：
0
作者：
Jex AR;Svärd S;Hagen KD;Starcevich H;Emery-Corbin SJ;Balan B;Nosala C;Dawson SC
通讯作者：
Dawson SC

Robust and stable transcriptional repression in Giardia using CRISPRi.

DOI：
10.1091/mbc.e18-09-0605
发表时间：
2019-01-01
期刊：
Molecular biology of the cell
影响因子：
3.3
作者：
McInally SG;Hagen KD;Nosala C;Williams J;Nguyen K;Booker J;Jones K;Dawson SC
通讯作者：
Dawson SC