VAP-A and its binding partner CERT drive biogenesis of RNA-containing extracellular vesicles at ER membrane contact sites.
VAP-A and its binding partner CERT drive biogenesis of RNA-containing extracellular vesicles at ER membrane contact sites.
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DOI：
10.1016/j.devcel.2022.03.012
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发表时间：
2022-04-25
影响因子：
11.8
通讯作者：
Weaver, Alissa M.
中科院分区：
文献类型：
--
作者：
Barman, Bahnisikha;Sung, Bong Hwan;Krystofiak, Evan;Ping, Jie;Ramirez, Marisol;Millis, Bryan;Allen, Ryan;Prasad, Nripesh;Chetyrkin, Sergei;Calcutt, M. Wade;Vickers, Kasey;Patton, James G.;Liu, Qi;Weaver, Alissa M.
通过细胞外囊泡（ev）的RNA转移影响细胞表型；然而，缺乏关于含rna的电动汽车的生物发生的信息限制了该领域的进展。在这里，我们确定内质网膜接触位点（ER MCS）是产生含有rna的ev的平台。我们确定了一个小ev亚群，其RNA高度富集，并由ER MCS连接蛋白VAP-A调节。在功能上，vap - a调节的ev对miR-100在细胞间转移和体内肿瘤形成至关重要。脂质分析显示，vap - a敲低的EV生物源性脂质神经酰胺减少。与vap - a结合的神经酰胺转移蛋白CERT的敲低导致了类似的EV RNA含量缺陷。影像学实验显示，VAP-A促进多泡体（multivesicular bodies, MVBs）的腔内填充，CERT定位于MVBs，神经酰胺生成酶中性鞘磷脂酶2与VAP-A阳性ER共定位。我们提出通过VAP-A-CERT连接的神经酰胺转移驱动选定的含有rna的EV群体的生物发生。含rna的细胞外囊泡（ev）的生物发生机制尚不清楚。Barman等人描述了内质网膜接触位点（ER MCS）、ER MCS连接体VAP-A和神经酰胺转运体CERT对含有rna的ev的亚群的生物发生至关重要的途径。
RNA transfer via extracellular vesicles (EVs) influences cell phenotypes; however, lack of information regarding biogenesis of RNA-containing EVs has limited progress in the field. Here, we identify endoplasmic reticulum membrane contact sites (ER MCS) as platforms for generation of RNA-containing EVs. We identify a subpopulation of small EVs that is highly enriched in RNA and regulated by the ER MCS linker protein VAP-A. Functionally, VAP-A-regulated EVs are critical for miR-100 transfer between cells and in vivo tumor formation. Lipid analysis of VAP-A-knockdown EVs revealed reductions in the EV biogenesis lipid ceramide. Knockdown of the VAP-A-binding ceramide transfer protein CERT led to similar defects in EV RNA content. Imaging experiments revealed that VAP-A promotes luminal filling of multivesicular bodies (MVBs), CERT localizes to MVBs, and the ceramide-generating enzyme neutral sphingomyelinase 2 colocalizes with VAP-A-positive ER. We propose that ceramide transfer via VAP-A-CERT linkages drives biogenesis of a select RNA-containing EV population. Biogenesis of RNA-containing extracellular vesicles (EVs) is poorly understood. Barman et al. delineate a pathway where endoplasmic reticulum membrane contact sites (ER MCS), the ER MCS linker VAP-A, and the ceramide transporter CERT are essential for biogenesis of a select subpopulation of RNA-containing EVs.
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影响因子：
3.5
作者：
De Vos KJ;Mórotz GM;Stoica R;Tudor EL;Lau KF;Ackerley S;Warley A;Shaw CE;Miller CC
通讯作者：
Miller CC
影响因子：
11.8
作者：
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16
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Lötvall J
影响因子：
14.9
作者：
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通讯作者：
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影响因子：
8.8
作者：
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通讯作者：
Ansel, K. Mark