A human polymorphism affects NEDD4L subcellular targeting by leading to two isoforms that contain or lack a C2 domain.
A human polymorphism affects NEDD4L subcellular targeting by leading to two isoforms that contain or lack a C2 domain.
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DOI：
10.1186/1471-2121-10-26
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发表时间：
2009-04-13
期刊：
影响因子：
--
通讯作者：
Rohrwasser A
中科院分区：
文献类型：
--
作者：
Garrone NF;Blazer-Yost BL;Weiss RB;Lalouel JM;Rohrwasser A
泛素化具有多种细胞功能，包括蛋白酶体降解以及对多种蛋白质的稳定性、功能和细胞内定位的控制。 NEDD4L 是 E3 泛素连接酶 HECT 类的成员。 NEDD4L 蛋白亚型的一个决定性特征是存在或不存在氨基末端 C2 结构域，这是一类亚细胞钙依赖性靶向结构域。我们之前在人类 NEDD4L 中发现了一个常见的变体，它产生包含或缺乏 C2 结构域的亚型。为了解决 NEDD4L 常见变体对 NEDD4L 亚细胞定位的潜在功能意义，用增强型绿色荧光蛋白标记包含或缺乏 C2 结构域的 NEDD4L 亚型，转染到非洲爪蟾肾上皮细胞中，并通过对活细胞进行共聚焦显微镜成像。我们报告，该 C2 结构域的存在或缺失对 NEDD4L 的亚细胞分布、含有和缺乏 NEDD4L 同工型的 C2 响应钙刺激的能力以及 NEDD4L 底物 ENaC 亚基的细胞内转运产生不同的影响。此外，含 C2 的异构体影响 β-ENaC 从细胞内库动员的能力涉及 NEDD4L 泛素化活性位点。我们提出了一个模型来解释这种常见遗传变异在细胞水平上对蛋白质功能的潜在影响。包含或缺乏 C2 结构域的 NEDD4L 亚型针对不同的细胞内位置。此外，含有 C2 的 NEDD4L 同工型能够响应钙刺激而在质膜和细胞内区室之间穿梭，而缺乏 C2 的同工型则不能。含有 C2 的亚型对细胞内池中 ENaC 亚基的动员有不同的影响，并且该运输步骤需要 NEDD4L 泛素连接酶活性。这一观察结果表明 cAMP 介导的 ENaC 胞吐作用中需要 PY 基序的新机制。我们已经阐明了常见的遗传变异如何在细胞水平上构成 NEDD4L 显着功能多样性的基础。我们提出了一个模型来描述功能变化如何影响血压。此外，我们对 NEDD4L 亚型差异功能的观察可能会影响涉及这种泛素连接酶的生理学的其他方面。
Ubiquitination serves multiple cellular functions, including proteasomal degradation and the control of stability, function, and intracellular localization of a wide variety of proteins. NEDD4L is a member of the HECT class of E3 ubiquitin ligases. A defining feature of NEDD4L protein isoforms is the presence or absence of an amino-terminal C2 domain, a class of subcellular, calcium-dependent targeting domains. We previously identified a common variant in human NEDD4L that generates isoforms that contain or lack a C2 domain. To address the potential functional significance of the NEDD4L common variant on NEDD4L subcellular localization, NEDD4L isoforms that either contained or lacked a C2 domain were tagged with enhanced green fluorescent protein, transfected into Xenopus laevis kidney epithelial cells, and imaged by performing confocal microscopy on live cells. We report that the presence or absence of this C2 domain exerts differential effects on the subcellular distribution of NEDD4L, the ability of C2 containing and lacking NEDD4L isoforms to mobilize in response to a calcium stimulus, and the intracellular transport of subunits of the NEDD4L substrate, ENaC. Furthermore, the ability of the C2-containing isoform to influence β-ENaC mobilization from intracellular pools involves the NEDD4L active site for ubiquitination. We propose a model to account for the potential impact of this common genetic variant on protein function at the cellular level. NEDD4L isoforms that contain or lack a C2 domain target different intracellular locations. Additionally, whereas the C2-containing NEDD4L isoform is capable of shuttling between the plasma membrane and intracellular compartments in response to calcium stimulus the C2-lacking isoform can not. The C2-containing isoform differentially affects the mobilization of ENaC subunits from intracellular pools and this trafficking step requires NEDD4L ubiquitin ligase activity. This observation suggests a new mechanism for the requirement for the PY motif in cAMP-mediated exocytosis of ENaC. We have elucidated how a common genetic variant can underlie significant functional diversity in NEDD4L at the cellular level. We propose a model that describes how that functional variation may influence blood pressure. Moreover, our observations regarding differential function of the NEDD4L isoforms may impact other aspects of physiology that involve this ubiquitin ligase.
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NEDD4L中的多态性与盐敏感性升高，P-肾脏水平降低以及NT-ProANP水平增加有关。
影响因子：
3.7
作者：
Dahlberg, Jonas;Nilsson, Lars-Olof;von Wowern, Fredrik;Melander, Olle
通讯作者：
Melander, Olle
影响因子：
2.3
作者：
Hollenberg, NK
通讯作者：
Hollenberg, NK
影响因子：
30.8
作者：
HANSSON, JH;NELSONWILLIAMS, C;LIFTON, RP
通讯作者：
LIFTON, RP
DOI：
10.1152/ajprenal.2000.279.1.f46
发表时间：
2000-07-01
影响因子：
4.2
作者：
Ecelbarger, CA;Kim, GH;Knepper, MA
通讯作者：
Knepper, MA
影响因子：
19.6
作者：
Fava, C.;von Wowern, F.;Melander, O.
通讯作者：
Melander, O.