A novel motif in the proximal C-terminus of Pannexin 1 regulates cell surface localization
A novel motif in the proximal C-terminus of Pannexin 1 regulates cell surface localization
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DOI：
10.1038/s41598-019-46144-5
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发表时间：
2019-07-05
影响因子：
4.6
通讯作者：
Swayne, Leigh Anne
中科院分区：
文献类型：
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作者：
Epp, Anna L.;Ebert, Sarah N.;Swayne, Leigh Anne
Pannexin 1（Panx 1）离子和代谢物通道在多种细胞中表达，它调节许多细胞行为，包括增殖和分化。Panx 1在细胞表面以及细胞内膜上表达。以前的工作表明，近端Panx 1 C-末端（Panx 1CT）内的区域调节细胞表面定位。在这里，我们报告了一个假定的富含亮氨酸的重复序列（LRR）基序的发现在近端Panx 1CT必要的Panx 1细胞表面表达在HEK 293 T细胞。假定的LRR基序的缺失导致Panx 1细胞表面分布的显著损失。互补细胞表面寡聚化和糖基化状态分析的结果与Panx 1 LRR缺失突变体的细胞表面表达降低一致。值得注意的是，寡聚化分析揭示了在细胞表面存在假定的Panx 1二聚体和三聚体。Panx 1的表达增加了HEK 293 T细胞的生长并减少了倍增时间，而Panx 1 LRR缺失突变体（高度保守的片段）的表达没有再现这种效果。总之，在这里，我们发现了一个假定的LRR基序在Panx 1CT的影响Panx 1细胞表面定位的存在。总的来说，这些发现提供了新的见解的C-末端调控的Panx 1贩运的分子机制，并提出了潜在的新的调查方面的Panx 1寡聚化和糖基化。
The Pannexin 1 (Panx1) ion and metabolite channel is expressed in a wide variety of cells where it regulates a number of cell behaviours including proliferation and differentiation. Panx1 is expressed on the cell surface as well as intracellular membranes. Previous work suggests that a region within the proximal Panx1 C-terminus (Panx1CT) regulates cell surface localization. Here we report the discovery of a putative leucine-rich repeat (LRR) motif in the proximal Panx1CT necessary for Panx1 cell surface expression in HEK293T cells. Deletion of the putative LRR motif results in significant loss of Panx1 cell surface distribution. Outcomes of complementary cell surface oligomerization and glycosylation state analyses were consistent with reduced cell surface expression of Panx1 LRR deletion mutants. Of note, the oligomerization analysis revealed the presence of putative dimers and trimers of Panx1 at the cell surface. Expression of Panx1 increased HEK293T cell growth and reduced doubling time, while expression of a Panx1 LRR deletion mutant (highly conserved segment) did not reproduce this effect. In summary, here we discovered the presence of a putative LRR motif in the Panx1CT that impacts on Panx1 cell surface localization. Overall these findings provide new insights into the molecular mechanisms underlying C-terminal regulation of Panx1 trafficking and raise potential new lines of investigation with respect to Panx1 oligomerization and glycosylation.