Elucidating the role of SNAREs in membrane contact site formation and cholesterol homeostasis

阐明 SNARE 在膜接触位点形成和胆固醇稳态中的作用

• 批准号: BB/S009566/1
• 负责人: Andrew Peden
• 金额: $ 72.32万
• 依托单位: University of Sheffield
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FS009566%2F1
• 关键词: Elucidating; role; SNAREs; membrane; contact

The human body is made of billions of cells and each cell is surrounded by a membrane made from lipids and proteins. In addition, each cell also contains numerous specialised compartments called organelles, which are also surrounded by membranes. A key lipid found in many membranes is cholesterol. Cholesterol can be obtained from the food that we eat or is made in a cellular compartment called the endoplasmic reticulum. Imbalances in cholesterol levels are toxic to cells so its levels are tightly regulated. However, this process can go wrong and defects in cholesterol regulation are associated with a range of diseases from atherosclerosis through to neurodegeneration. The aim of this research proposal is to work out how cholesterol is transported within cells and determine how this process regulates cholesterol levels throughout the body. Preliminary work performed by our research team has identified that a protein called VAMP4 is important for this process. When we disrupt the function of VAMP4 in cells and in mice we see that the levels of cholesterol are significantly altered. Using a technique called electron microscopy we can directly look at organelles within cells and see how they interact with each other. In cells lacking VAMP4, we can see that an organelle called an endosome is not properly bound to the endoplasmic reticulum. In addition, when cells are made to make too much VAMP4 we see that more of the endoplasmic reticulum is bound to endosomes. Taken all of these results in consideration, it is our current model that VAMP4 is working to bring endosomes and the endoplasmic reticulum in very close proximity, which then allows cholesterol to move between these organelles. At present, it is unclear how this process is regulated so we plan to use a range of biochemical techniques, including mass spectrometry to identify which proteins interact with VAMP4 and co-ordinate this process. To determine how loss of VAMP4 leads to global changes in cholesterol levels we plan to investigate the function of VAMP4 in macrophages, which is a cell type with an important role in cholesterol biology. In summary, the research outlined in this proposal will increase our understanding of important cellular pathways, which regulate cholesterol levels and in the long term provide a foundation for understanding how nutrition and ageing effect cholesterol physiology.

人体由数十亿个细胞组成，每个细胞都被脂质和蛋白质组成的膜包围。此外，每个细胞还包含许多称为细胞器的专门隔室，这些隔室也被膜包围。许多细胞膜中发现的一种关键脂质是胆固醇。胆固醇可以从我们吃的食物中获得，或者在称为内质网的细胞隔室中产生。胆固醇水平的不平衡对细胞是有毒的，所以它的水平受到严格的控制。然而，这一过程可能出错，胆固醇调节的缺陷与从动脉粥样硬化到神经变性的一系列疾病有关。这项研究计划的目的是找出胆固醇如何在细胞内运输，并确定这一过程如何调节整个身体的胆固醇水平。我们的研究小组进行的初步工作已经确定，一种名为VAMP4的蛋白质对这一过程很重要。当我们在细胞和小鼠中破坏VAMP4的功能时，我们看到胆固醇的水平发生了显着变化。使用一种称为电子显微镜的技术，我们可以直接观察细胞内的细胞器，并观察它们如何相互作用。在缺乏VAMP4的细胞中，我们可以看到一种叫做内体的细胞器没有正确地结合到内质网上。此外，当细胞产生过多的VAMP4时，我们看到更多的内质网与核内体结合。考虑到所有这些结果，我们目前的模型是VAMP4正在努力使内体和内质网非常接近，然后允许胆固醇在这些细胞器之间移动。目前，还不清楚这一过程是如何调节的，因此我们计划使用一系列生化技术，包括质谱法来鉴定哪些蛋白质与VAMP4相互作用并协调这一过程。为了确定VAMP4的丢失如何导致胆固醇水平的整体变化，我们计划研究VAMP4在巨噬细胞中的功能，巨噬细胞是一种在胆固醇生物学中具有重要作用的细胞类型。总之，本提案中概述的研究将增加我们对调节胆固醇水平的重要细胞途径的理解，并从长远来看为理解营养和衰老如何影响胆固醇生理学提供基础。

项目成果

Organelle tethering, pore formation and SNARE compensation in the late endocytic pathway.

• DOI: 10.1242/jcs.255463
• 发表时间: 2021-05-15
• 期刊: Journal of cell science
• 影响因子: 4
• 作者: Davis LJ;Bright NA;Edgar JR;Parkinson MDJ;Wartosch L;Mantell J;Peden AA;Luzio JP
• 通讯作者: Luzio JP

Cholesterol Overload: Contact Sites to the Rescue!

• DOI: 10.1177/2515256419893507
• 发表时间: 2019-01-01
• 期刊: Contact (Thousand Oaks (Ventura County, Calif.))
• 作者: Enrich, Carlos;Rentero, Carles;Eden, Emily R
• 通讯作者: Eden, Emily R

An Open-Source Framework for Automated High-Throughput Cell Biology Experiments.

• DOI: 10.3389/fcell.2021.697584
• 发表时间: 2021
• 期刊: Frontiers in cell and developmental biology
• 影响因子: 5.5
• 作者: Katunin P;Zhou J;Shehata OM;Peden AA;Cadby A;Nikolaev A
• 通讯作者: Nikolaev A

Staying in touch with the endocytic network: The importance of contacts for cholesterol transport.

• DOI: 10.1111/tra.12726
• 发表时间: 2020-05
• 期刊: Traffic (Copenhagen, Denmark)
• 作者: Martello A;Platt FM;Eden ER
• 通讯作者: Eden ER

Comparative host-coronavirus protein interaction networks reveal pan-viral disease mechanisms.

• DOI: 10.1126/science.abe9403
• 发表时间: 2020-12-04
• 期刊: Science (New York, N.Y.)
• 作者: Gordon DE;Hiatt J;Bouhaddou M;Rezelj VV;Ulferts S;Braberg H;Jureka AS;Obernier K;Guo JZ;Batra J;Kaake RM;Weckstein AR;Owens TW;Gupta M;Pourmal S;Titus EW;Cakir M;Soucheray M;McGregor M;Cakir Z;Jang G;O'Meara MJ;Tummino TA;Zhang Z;Foussard H;Rojc A;Zhou Y;Kuchenov D;Hüttenhain R;Xu J;Eckhardt M;Swaney DL;Fabius JM;Ummadi M;Tutuncuoglu B;Rathore U;Modak M;Haas P;Haas KM;Naing ZZC;Pulido EH;Shi Y;Barrio-Hernandez I;Memon D;Petsalaki E;Dunham A;Marrero MC;Burke D;Koh C;Vallet T;Silvas JA;Azumaya CM;Billesbølle C;Brilot AF;Campbell MG;Diallo A;Dickinson MS;Diwanji D;Herrera N;Hoppe N;Kratochvil HT;Liu Y;Merz GE;Moritz M;Nguyen HC;Nowotny C;Puchades C;Rizo AN;Schulze-Gahmen U;Smith AM;Sun M;Young ID;Zhao J;Asarnow D;Biel J;Bowen A;Braxton JR;Chen J;Chio CM;Chio US;Deshpande I;Doan L;Faust B;Flores S;Jin M;Kim K;Lam VL;Li F;Li J;Li YL;Li Y;Liu X;Lo M;Lopez KE;Melo AA;Moss FR 3rd;Nguyen P;Paulino J;Pawar KI;Peters JK;Pospiech TH Jr;Safari M;Sangwan S;Schaefer K;Thomas PV;Thwin AC;Trenker R;Tse E;Tsui TKM;Wang F;Whitis N;Yu Z;Zhang K;Zhang Y;Zhou F;Saltzberg D;QCRG Structural Biology Consortium;Hodder AJ;Shun-Shion AS;Williams DM;White KM;Rosales R;Kehrer T;Miorin L;Moreno E;Patel AH;Rihn S;Khalid MM;Vallejo-Gracia A;Fozouni P;Simoneau CR;Roth TL;Wu D;Karim MA;Ghoussaini M;Dunham I;Berardi F;Weigang S;Chazal M;Park J;Logue J;McGrath M;Weston S;Haupt R;Hastie CJ;Elliott M;Brown F;Burness KA;Reid E;Dorward M;Johnson C;Wilkinson SG;Geyer A;Giesel DM;Baillie C;Raggett S;Leech H;Toth R;Goodman N;Keough KC;Lind AL;Zoonomia Consortium;Klesh RJ;Hemphill KR;Carlson-Stevermer J;Oki J;Holden K;Maures T;Pollard KS;Sali A;Agard DA;Cheng Y;Fraser JS;Frost A;Jura N;Kortemme T;Manglik A;Southworth DR;Stroud RM;Alessi DR;Davies P;Frieman MB;Ideker T;Abate C;Jouvenet N;Kochs G;Shoichet B;Ott M;Palmarini M;Shokat KM;García-Sastre A;Rassen JA;Grosse R;Rosenberg OS;Verba KA;Basler CF;Vignuzzi M;Peden AA;Beltrao P;Krogan NJ
• 通讯作者: Krogan NJ