Subcellular localization and function of alternatively spliced Noxo1 isoforms
Subcellular localization and function of alternatively spliced Noxo1 isoforms
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DOI：
10.1016/j.freeradbiomed.2006.08.024
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发表时间：
2007-01-15
影响因子：
7.4
通讯作者：
Leto, Thomas L.
中科院分区：
文献类型：
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作者：
Ueyama, Takehiko;Lekstrom, Kristen;Leto, Thomas L.
Nox组织者1 （Noxo1）是p47（phox）同源物，通过选择性mRNA剪接产生具有独特n端PX结构域的四个同种异构体。我们比较了这些异构体的亚细胞分布或它们作为GFP融合蛋白产生的分离的PX结构域，以及它们在几种转染模型中支持Nox1活性的能力。Noxo1 α、β、γ和δ表现出不同的亚细胞定位模式，这是由它们的PX结构域决定的。在HEK293细胞中，Noxo1 β表现出明显的质膜结合，Noxo1 γ表现出质膜和核结合，Noxo1 α和Noxo1 6主要定位于细胞内囊泡或细胞质聚集体，而不是质膜。在HEK293细胞中，Noxo1活性与Noxo1质膜结合相关，因为Noxo1 β支持最高活性，而Noxo1 γ和Noxo1 α分别支持中等或低活性。在nos -7细胞中，Noxo1 α定位于质膜上，三种同工异构体（α、β和γ）支持的活性没有显著差异。和的PX结构域结合相同的磷脂，包括磷脂酸。这些结果表明，变异的PX结构域是Noxo1定位和Nox1功能的唯一决定因素。最后，尽管将p22（phox）运输到质膜需要Nox1，但过表达的Noxo1亚型并不影响p22（phox）的定位。Elsevier Inc.出版。
Nox organizer 1 (Noxo1), a p47(phox) homolog, is produced as four isoforms with unique N-terminal PX domains derived by alternative mRNA splicing. We compared the subcellular distribution of these isoforms or their isolated PX domains produced as GFP fusion proteins, as well as their ability to support Nox1 activity in several transfected models. Noxo1 alpha, beta, gamma, and delta show different subcellular localization patterns, determined by their PX domains. In HEK293 cells, Noxo1 beta exhibits prominent plasma membrane binding, Noxo1 gamma shows plasma membrane and nuclear associations, and Noxo1 alpha and 6 localize primarily on intracellular vesicles or cytoplasmic aggregates, but not the plasma membrane. Nox1 activity correlates with Noxo1 plasma membrane binding in HEK293 cells, since Noxo1 beta supports the highest activity and Noxo1 gamma and Noxo1 alpha support moderate or low activities, respectively. In COS-7 cells, where Noxo1 alpha localizes on the plasma membrane, the activities supported by the three isoforms (alpha, beta, and gamma) do not differ significantly. The PX domains of beta and gamma bind the same phospholipids, including phosphatidic acid. These results indicate that the variant PX domains are unique determinants of Noxo1 localization and Nox1 function. Finally, the overexpressed Noxo1 isoforms do not affect p22(phox) localization, although Nox1 is needed to transport p22(phox) to the plasma membrane. Published by Elsevier Inc.