Human gut bacteria tailor extracellular vesicle cargo for the breakdown of diet- and host-derived glycans.
Human gut bacteria tailor extracellular vesicle cargo for the breakdown of diet- and host-derived glycans.
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DOI：
10.1073/pnas.2306314120
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发表时间：
2023-07-04
影响因子：
11.1
通讯作者：
Feldman, Mario F.
中科院分区：
文献类型：
--
作者：
Sartorio, Mariana G.;Pardue, Evan J.;Scott, Nichollas E.;Feldman, Mario F.
类杆菌是人类肠道微生物区系中的一个重要属，能产生大量携带特定蛋白质的OMV(外膜小泡)。在这里，我们证明了蛋白质在OMV中的分类依赖于一个带负电荷的“脂蛋白输出信号”(LES)。我们利用外膜标记和OMV标记融合到荧光蛋白上，通过时间推移荧光显微镜显示OMV的生物发生。此外，我们还进行了比较蛋白质组学分析，结果表明Bt(Bacteroids Thetaiotaomicron)可以主动改变OMV的含量，从而优化多糖的利用。我们得出结论，OMV的生产在Bt中受到高度调控。尽管OMV在类杆菌属中的作用已得到公认，但对其生物发生机制知之甚少。未来的研究将集中于进一步阐明这一未被充分描述的过程。细胞外小泡在生命的三个领域都有产生，它们的生物发生在真核生物和古生物中有着共同的古代起源。尽管细菌小泡在几十年前就被发现，并被认为具有多种作用，但细菌中小泡的生物发生机制尚未建立。出于这个原因，人们对细菌小泡的生物学相关性有很大程度的怀疑。类杆菌是人类肠道微生物区系中的一个重要成员，它能产生大量的外膜小泡(OMV)，而外膜囊泡被认为具有重要的生理作用。在这里，我们使用了一个双标记系统，包括外膜和OMV特异的标记融合到荧光蛋白上，通过时间推移显微镜观察OMV的生物发生。此外，我们进行了比较蛋白质组学分析，表明在Bt中，OMV货物适合于不同多糖的最佳利用。我们还表明，带负电荷的N-末端基序作为蛋白质分选到OMV的信号，与营养物质的可用性无关。我们的结果表明，OMV的产生是Bt基因高度调控的过程的结果。
Bacteroides is a prominent genus of the human gut microbiota, which produces large amounts of OMVs (outer membrane vesicles) carrying specific protein cargo. Here, we show that sorting of proteins into OMVs depends on a negatively charged “lipoprotein export signal” (LES). We employed OM (outer membrane) and OMV markers fused to fluorescent proteins to show OMV biogenesis via time-lapse fluorescence microscopy. Additionally, we performed comparative proteomic analyses showing that Bt (Bacteroides thetaiotaomicron) actively alters the OMV content to optimize the utilization of polysaccharides. We conclude that OMV production is highly regulated in Bt. Despite the recognized roles of OMVs in Bacteroides, little is known about their biogenesis. Future studies will focus on further elucidating this undercharacterized process. Extracellular vesicles are produced in all three domains of life, and their biogenesis has common ancient origins in eukaryotes and archaea. Although bacterial vesicles were discovered several decades ago and multiple roles have been attributed to them, no mechanism has been established for vesicles biogenesis in bacteria. For this reason, there is a significant level of skepticism about the biological relevance of bacterial vesicles. Bacteroides thetaiotaomicron (Bt), a prominent member of the human intestinal microbiota, produces significant amounts of outer membrane vesicles (OMVs) which have been proposed to play key physiological roles. Here, we employed a dual marker system, consisting of outer membrane- and OMV-specific markers fused to fluorescent proteins to visualize OMV biogenesis by time-lapse microscopy. Furthermore, we performed comparative proteomic analyses to show that, in Bt, the OMV cargo is adapted for the optimal utilization of different polysaccharides. We also show that a negatively charged N-terminal motif acts as a signal for protein sorting into OMVs irrespective of the nutrient availability. Our results demonstrate that OMV production is the result of a highly regulated process in Bt.
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影响因子：
30.3
作者：
Martens EC;Chiang HC;Gordon JI
通讯作者：
Gordon JI
DOI：
10.1073/pnas.2009556117
发表时间：
2020-09-29
影响因子：
11.1
作者：
Garcia-Bayona, Leonor;Coyne, Michael J.;Comstock, Laurie E.
通讯作者：
Comstock, Laurie E.
影响因子：
6.4
作者：
Elhenawy W;Debelyy MO;Feldman MF
通讯作者：
Feldman MF
影响因子：
64.8
作者：
Rakoff-Nahoum S;Foster KR;Comstock LE
通讯作者：
Comstock LE
影响因子：
9.8
作者：
Martens EC;Lowe EC;Chiang H;Pudlo NA;Wu M;McNulty NP;Abbott DW;Henrissat B;Gilbert HJ;Bolam DN;Gordon JI
通讯作者：
Gordon JI