Hu Yang, Chuang Jen-Zen, Xu Kai, McGraw Timothy G, Sung Ching-Hwa
Graduate Program of Neuroscience, Weill Medical College of Cornell University, 1300 York Avenue, New York, NY 10021, USA.
J Cell Sci. 2002 Dec 15;115(Pt 24):4755-63. doi: 10.1242/jcs.00177.
Rab5, a member of the small GTPase family of proteins, is primarily localized on early endosomes and has been proposed to participate in the regulation of early endosome trafficking. It has been reported that phosphatidylinositol 3-kinases and FYVE domain proteins, such as EEA1, can be recruited onto early endosomes and act as Rab5 effectors. SARA (Smad anchor for receptor activation), also a FYVE domain protein, was initially isolated as a participant in signal transduction from the transforming growth factor beta receptor. Overexpressed SARA has been found on EEA1-positive early endosomes. In this report, we show that endogenous SARA is present on early endosomes and overexpression of SARA causes endosomal enlargement. Functionally, SARA overexpression significantly delays the recycling of transferrin. The transferrin receptor distributed on the cell surfaces was also greatly reduced in cells overexpressing SARA. However, the internalization rate of transferrin is not affected by SARA overexpression. The morphological and functional alterations caused by SARA overexpression resemble those caused by overexpression of Rab5:GTP mutant Rab5Q79L. Finally, all SARA-mediated phenotypic changes can be counteracted by overexpression Rab5:GDP mutant Rab5S34N. These results collectively suggested that SARA plays an important functional role downstream of Rab5-regulated endosomal trafficking.
Rab5是小GTPase蛋白家族的成员之一,主要定位于早期内体,并被认为参与早期内体运输的调节。据报道,磷脂酰肌醇3激酶和含FYVE结构域的蛋白,如EEA1,可以被招募到早期内体上,并作为Rab5效应器发挥作用。SARA(受体激活的Smad锚定蛋白)也是一种含FYVE结构域的蛋白,最初作为转化生长因子β受体信号转导的参与者被分离出来。已发现在EEA1阳性的早期内体上有过表达的SARA。在本报告中,我们表明内源性SARA存在于早期内体上,且SARA的过表达会导致内体增大。在功能上,SARA的过表达显著延迟了转铁蛋白的循环利用。在过表达SARA的细胞中,分布在细胞表面的转铁蛋白受体也大大减少。然而,转铁蛋白的内化速率不受SARA过表达的影响。SARA过表达引起的形态和功能改变类似于Rab5的GTP突变体Rab5Q79L过表达所引起的改变。最后,所有SARA介导的表型变化都可以通过过表达Rab5的GDP突变体Rab5S34N来抵消。这些结果共同表明,SARA在Rab5调节的内体运输下游发挥重要的功能作用。