Identification of the Type IX Secretion System Component, PorV (CHU_3238), Involved in Secretion and Localization of Proteins in Cytophaga hutchinsonii.
Identification of the Type IX Secretion System Component, PorV (CHU_3238), Involved in Secretion and Localization of Proteins in Cytophaga hutchinsonii.
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鉴定参与哈钦森噬细胞菌蛋白质分泌和定位的 IX 型分泌系统组件 PorV (CHU_3238)
DOI：
10.3389/fmicb.2021.742673
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发表时间：
2021
影响因子：
5.2
通讯作者：
Lu X
中科院分区：
文献类型：
--
作者：
Zhao D;Song W;Wang S;Zhang W;Zhao Y;Lu X
Cytophaga hutchinsonii可以有效地降解纤维素并在表面上快速滑行，但其潜在机制尚不清楚。IX型分泌系统（T9 SS）参与蛋白质分泌和滑行运动，这是拟杆菌门所特有的。在这项研究中，我们删除了一个同源基因的PorV（chu_3238），穿梭蛋白在T9 SS。Δ3238突变体导致纤维素分解和滑动缺陷，而其他拟杆菌中的porV缺失突变体可以正常滑动。添加Ca ~（2+）和K ~+促进了PY 6培养基中的生长，表明chu_3238在离子吸收中的潜在作用。蛋白质组学分析显示，与野生型（WT）相比，Δ3238突变体中细胞外蛋白的数量增加，外膜蛋白减少。与WT相比，Δ3238完整细胞中的内切葡聚糖酶活性降低了约70%。这些结果表明，在Δ3238突变体中，分泌的蛋白不能附着到细胞表面，而是释放到细胞外空间中。然而，货物蛋白积累在周质的其他报告porV缺失突变体。此外，在3238-FLAG菌株中，通过共免疫沉淀将易位子SprA和Plug蛋白的同源物拉下，其参与约氏黄杆菌的T9 SS中的蛋白质转运。在Δ3238突变体中，脂多糖（LPS）的完整性也受到影响，这可能是细胞对毒性试剂敏感的原因。CHU_3238的功能多样性表明其在C. hutchinsonii的研究，并强调了拟杆菌中PorV在T9 SS中的功能差异。
Cytophaga hutchinsonii can efficiently degrade cellulose and rapidly glide over surfaces, but the underlying mechanisms remain unclear. The type IX secretion system (T9SS) is involved in protein secretion and gliding motility, which is unique to the phylum Bacteroidetes. In this study, we deleted a homologous gene of PorV (chu_3238), a shuttle protein in the T9SS. The Δ3238 mutant caused cellulolytic and gliding defects, while the porV deletion mutants in other Bacteroidetes could glide normally. Adding Ca2+ and K+ improved growth in the PY6 medium, suggesting a potential role of chu_3238 in ion uptake. A proteomic analysis showed an increase in the number of extracellular proteins in the Δ3238 mutant and a decrease in the outer membrane proteins compared to the wild type (WT). Endoglucanase activity in the Δ3238 intact cells was reduced by approximately 70% compared to that of the WT. These results indicate that the secreted proteins could not attach to the cell surface but were released into the extracellular space in the Δ3238 mutant. However, the cargo proteins accumulated in the periplasm of other reported porV deletion mutants. In addition, the homologs of the translocon SprA and a Plug protein were pulled down by co-immunoprecipitation in the 3238-FLAG strain, which are involved in protein transport in the T9SS of Flavobacterium johnsoniae. The integrity of the lipopolysaccharide (LPS) was also affected in the Δ3238 mutant, which may be the reason for the sensitivity of the cell to toxic reagents. The functional diversity of CHU_3238 suggests its important role in the T9SS of C. hutchinsonii and highlights the functional differences of PorV in the T9SS among the Bacteroidetes.
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Cytophaga hutchinsonii SprA 和 SprT 是 Ca2 获取、纤维素降解和细胞运动所需的 IX 型分泌系统的重要组成部分
影响因子：
5.2
作者：
Gao L;Tan Y;Zhang W;Qi Q;Lu X
通讯作者：
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影响因子：
3.2
作者：
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影响因子：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
5.2
作者：
Pérez-Pascual D;Rochat T;Kerouault B;Gómez E;Neulat-Ripoll F;Henry C;Quillet E;Guijarro JA;Bernardet JF;Duchaud E
通讯作者：
Duchaud E