Junctional Localization of Septin 2 Is Required for Organization of Junctional Proteins in Static Endothelial Monolayers.
Junctional Localization of Septin 2 Is Required for Organization of Junctional Proteins in Static Endothelial Monolayers.
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DOI：
10.1161/atvbaha.120.315472
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发表时间：
2021-01
期刊：
影响因子：
--
通讯作者：
Cooper JA
中科院分区：
文献类型：
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作者：
Kim J;Cooper JA
Septin 2 在人微血管内皮单层的连接处富集。 septin 2 的连接定位对于内皮细胞细胞间粘附蛋白的组织是必需的。通过使用 shRNA 抑制表达、用炎性细胞因子、TNF-α 处理以及与质膜相互作用的 Septin 2 PIP2 结合突变体 (BM) 缺陷的异位过度表达，Septin 2 在连接处被耗尽。在这些条件下，分析了各种连接蛋白的组织和表达水平。免疫荧光染色的共聚焦图像显示粘附连接蛋白、nectin-2 和 afadin、紧密连接蛋白、ZO-1 和细胞间粘附蛋白、PECAM-1 的严重破坏。除了 nectin-2 的表达大幅增加外，这些蛋白质的免疫印迹并未显示出显着的表达变化。通过 septin 2 抑制和使用 RNAseq 进行 TNF-α 处理进行的显着差异基因表达谱和生物途径分析显示出共同的重叠途径。表达的共性可能与细胞间粘附蛋白的整体组织的类似作用一致。 Septin 2 在细胞连接处的定位对于连接蛋白的排列和内皮单层形成的屏障的完整性是必需的。
Septin 2 is enriched at junctions in human microvascular endothelial monolayers. The junctional localization of septin 2 is necessary for organization of cell-cell adhesion proteins of endothelial cells. Septin 2 was depleted at jucntions by suppression of expression using shRNA, treatment with inflammatory cytokine, TNF-α, and ectopic overexpression of septin 2 PIP2 binding mutant (BM) defect in interaction with plasma membrane. Under those conditions, organizations and expression levels of various junctional proteins were analysed. Confocal images of immunofluorescence staining showed substantial disorganization of adherens junctional proteins, nectin-2 and afadin, tight junction protein, ZO-1, and intercellular adhesion protein, PECAM-1. Immunoblots for those proteins did not show significant changes in expression except for nectin-2 that highly increased in expression. Signifcant differential gene expression profiles and biological pathway analysis by septin 2 suppression and by TNF-α treatment using RNAseq showed common overlapping pathways. The commonalities in expression may be consistent with the similar effects on the overall organization of cell-cell adhesion proteins. Localization of septin 2 at cell junctions are required for the arrangement of junctional proteins and the integrity of the barrier formed by endothelial monolayers.
DOI：
10.1083/jcb.200507017
发表时间：
2005-12-05
期刊：
The Journal of cell biology
影响因子：
--
作者：
Gillis AN;Thomas S;Hansen SD;Kaplan KB
通讯作者：
Kaplan KB