A PI(3,5)P2 reporter reveals PIKfyve activity and dynamics on macropinosomes and phagosomes.
A PI(3,5)P2 reporter reveals PIKfyve activity and dynamics on macropinosomes and phagosomes.
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DOI：
10.1083/jcb.202209077
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发表时间：
2023-09-04
影响因子：
7.8
通讯作者：
King, Jason S.
中科院分区：
文献类型：
--
作者：
Vines, James H.;Maib, Hannes;Buckley, Catherine M.;Gueho, Aurelie;Zhu, Zhou;Soldati, Thierry;Murray, David H.;King, Jason S.
在这项工作中，作者描述了一种新的磷酸肌醇信号脂质PI（3，5）P2的生物传感器。他们证明，这一报告忠实地报告PI（3，5）P2的动态在Dictyosteobacteriumamoeopathy和哺乳动物细胞，并揭示了PI（3，5）P2的形成是如何在吞噬体成熟过程中调节的细节。磷脂酰肌醇信号脂质（PIP）是膜特性和运输的关键调节剂。其中，PI（3，5）P2是了解最少的之一，尽管在许多内吞途径（包括吞噬作用和巨胞饮作用）中起关键作用。PI（3，5）P2由磷酸肌醇5-激酶PIKfyve产生，其对于吞噬体消化和抗微生物活性至关重要。然而，由于缺乏可靠的报告基因，PI（3，5）P2动力学和调控仍然不清楚。使用变形虫Dictyosteelium discoideum，我们确定SnxA作为一个高度选择性的PI（3，5）P2结合蛋白，并表征其作为PI（3，5）P2在Dictyosteelium和哺乳动物细胞的报告。使用GFP-SnxA，我们表明，网骨藻吞噬体和macropinosomes积累PI（3，5）P2吞噬后3分钟，但随后保留不同，表明途径特异性调节。我们进一步发现PIKfyve募集和活性是可分离的，并且PIKfyve激活刺激其自身的解离。因此，SnxA是用于报告活细胞中PI（3，5）P2的新工具，其揭示了PIKfyve/PI（3，5）P2的作用和调节的关键机制细节。
In this work, the authors describe a new biosensor for the phosphoinositide signaling lipid PI(3,5)P2. They demonstrate that this reporter faithfully reports PI(3,5)P2 dynamics in both Dictyostelium amoebae and mammalian cells and uncover details of how PI(3,5)P2 formation is regulated during phagosome maturation. Phosphoinositide signaling lipids (PIPs) are key regulators of membrane identity and trafficking. Of these, PI(3,5)P2 is one of the least well-understood, despite key roles in many endocytic pathways including phagocytosis and macropinocytosis. PI(3,5)P2 is generated by the phosphoinositide 5-kinase PIKfyve, which is critical for phagosomal digestion and antimicrobial activity. However PI(3,5)P2 dynamics and regulation remain unclear due to lack of reliable reporters. Using the amoeba Dictyostelium discoideum, we identify SnxA as a highly selective PI(3,5)P2-binding protein and characterize its use as a reporter for PI(3,5)P2 in both Dictyostelium and mammalian cells. Using GFP-SnxA, we demonstrate that Dictyostelium phagosomes and macropinosomes accumulate PI(3,5)P2 3 min after engulfment but are then retained differently, indicating pathway-specific regulation. We further find that PIKfyve recruitment and activity are separable and that PIKfyve activation stimulates its own dissociation. SnxA is therefore a new tool for reporting PI(3,5)P2 in live cells that reveals key mechanistic details of the role and regulation of PIKfyve/PI(3,5)P2.
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影响因子：
16.6
作者：
Dong, Xian-ping;Shen, Dongbiao;Wang, Xiang;Dawson, Taylor;Li, Xinran;Zhang, Qi;Cheng, Xiping;Zhang, Yanling;Weisman, Lois S.;Delling, Markus;Xu, Haoxing
通讯作者：
Xu, Haoxing
DOI：
10.1111/j.1600-0854.2009.00915.x
发表时间：
2009-07
期刊：
Traffic (Copenhagen, Denmark)
影响因子：
--
作者：
de Lartigue J;Polson H;Feldman M;Shokat K;Tooze SA;Urbé S;Clague MJ
通讯作者：
Clague MJ
DOI：
10.1042/bj20090428
发表时间：
2009-07-29
期刊：
The Biochemical journal
影响因子：
--
作者：
Hammond GR;Schiavo G;Irvine RF
通讯作者：
Irvine RF
影响因子：
4
作者：
Berwick, DC;Dell, GC;Tavaré, JM
通讯作者：
Tavaré, JM
影响因子：
3.5
作者：
Hazeki, Kaoru;Nigorikawa, Kiyomi;Hazeki, Osamu
通讯作者：
Hazeki, Osamu