Identification of a novel mouse P4-ATPase family member highly expressed during spermatogenesis
Identification of a novel mouse P4-ATPase family member highly expressed during spermatogenesis
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鉴定在精子发生过程中高表达的新型小鼠 P4-ATPase 家族成员
DOI：
10.1242/jcs.047423
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发表时间：
2009-08
影响因子：
4
通讯作者：
Hanisch, Susanne
中科院分区：
文献类型：
--
作者：
Ding, Xiao-Yan;Okkeri, Juha;Xu, Peng;Hu, Rui-Ying;Xu, Qin;Pomorski, Thomas Guenther;Hanisch, Susanne
P4-ATP酶是真核生物特有的跨膜蛋白，在囊泡运输中起着重要作用。已提出它们作为磷脂翻转酶，从而调节细胞膜中的脂质拓扑结构。我们克隆并鉴定了一种新的小鼠睾丸特异性表达的P4-ATP酶，并将其命名为FetA（flippase expressed in testis splicing form A）。当在酿酒酵母中表达时，FetA部分定位于质膜，导致NBD标记的磷脂酰乙醇胺和磷脂酰胆碱的内化增加，支持FetA在跨细胞膜的向内脂质易位中的作用。在小鼠睾丸中，FetA蛋白存在于从粗线期精母细胞到成熟精子的配子细胞中，其细胞内定位与顶体形成密切相关，顶体形成是一个涉及密集的胞内囊泡形成和融合的过程。此外，在肥大细胞瘤P815细胞中通过RNA干扰FetA的功能丧失深刻地扰乱了高尔基复合体的结构组织，并导致在较低温度下组成性分泌的丧失。我们的研究结果表明，FetA在高尔基体形态和分泌功能中起着重要作用，这表明这种新型小鼠P4-ATP酶在精子发生中起着至关重要的作用。
P4-ATPases are transmembrane proteins unique to eukaryotes that play a fundamental role in vesicular transport. They have been proposed to act as phospholipid flippases thereby regulating lipid topology in cellular membranes. We cloned and characterized a novel murine P4-ATPase that is specifically expressed in testis, and named it FetA (flippase expressed in testis splicing form A). When expressed in Saccharomyces cerevisiae, FetA localizes partially to the plasma membrane resulting in increased internalization of NBD-labeled phosphatidylethanolamine and phosphatidylcholine, supporting a role for FetA in the inward lipid translocation across cellular membranes. In mouse testis, FetA protein is detected in gamete cells, from pachytene spermatocytes to mature sperms, and its intracellular localization is tightly related with acrosome formation, a process that involves intensive intracellular vesicle formation and fusion. Furthermore, loss-of-function of FetA by RNA interference in mastocytoma P815 cells profoundly perturbs the structural organization of the Golgi complex and causes loss of constitutive secretion at lower temperature. Our findings point to an essential role of FetA in Golgi morphology and secretory function, suggesting a crucial role for this novel murine P4-ATPase in spermatogenesis.
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