Sato Ken, Sato Miyuki, Audhya Anjon, Oegema Karen, Schweinsberg Peter, Grant Barth D
Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, NJ 08854, USA.
Mol Biol Cell. 2006 Jul;17(7):3085-94. doi: 10.1091/mbc.e06-03-0211. Epub 2006 May 3.
Caveolin is the major protein component required for the formation of caveolae on the plasma membrane. Here we show that trafficking of Caenorhabditis elegans caveolin-1 (CAV-1) is dynamically regulated during development of the germ line and embryo. In oocytes a CAV-1-green fluorescent protein (GFP) fusion protein is found on the plasma membrane and in large vesicles (CAV-1 bodies). After ovulation and fertilization the CAV-1 bodies fuse with the plasma membrane in a manner reminiscent of cortical granule exocytosis as described in other species. Fusion of CAV-1 bodies with the plasma membrane appears to be regulated by the advancing cell cycle, and not fertilization per se, because fusion can proceed in spe-9 fertilization mutants but is blocked by RNA interference-mediated knockdown of an anaphase-promoting complex component (EMB-27). After exocytosis, most CAV-1-GFP is rapidly endocytosed and degraded within one cell cycle. CAV-1 bodies in oocytes appear to be produced by the Golgi apparatus in an ARF-1-dependent, clathrin-independent, mechanism. Conversely endocytosis and degradation of CAV-1-GFP in embryos requires clathrin, dynamin, and RAB-5. Our results demonstrate that the distribution of CAV-1 is highly dynamic during development and provides new insights into the sorting mechanisms that regulate CAV-1 localization.
小窝蛋白是在质膜上形成小窝所需的主要蛋白质成分。在此我们表明,秀丽隐杆线虫小窝蛋白-1(CAV-1)的运输在生殖系和胚胎发育过程中受到动态调节。在卵母细胞中,一种CAV-1-绿色荧光蛋白(GFP)融合蛋白存在于质膜和大囊泡(CAV-1小体)中。排卵和受精后,CAV-1小体以一种类似于其他物种中描述的皮质颗粒胞吐作用的方式与质膜融合。CAV-1小体与质膜的融合似乎受细胞周期进程的调节,而非受精本身,因为融合可在spe-9受精突变体中进行,但会被RNA干扰介导的后期促进复合物成分(EMB-27)的敲低所阻断。胞吐作用后,大多数CAV-1-GFP在一个细胞周期内迅速被内吞并降解。卵母细胞中的CAV-1小体似乎是由高尔基体通过一种依赖ARF-1、不依赖网格蛋白的机制产生的。相反,胚胎中CAV-1-GFP的内吞作用和降解需要网格蛋白、发动蛋白和RAB-5。我们的结果表明,CAV-1的分布在发育过程中高度动态,并为调节CAV-1定位的分选机制提供了新的见解。