Characterization of Chlamydomonas reinhardtii zygote-specific cDNAs that encode novel proteins containing ankyrin repeats and WW domains.
Characterization of Chlamydomonas reinhardtii zygote-specific cDNAs that encode novel proteins containing ankyrin repeats and WW domains.
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DOI：
10.1104/pp.119.3.873
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发表时间：
1999-03
期刊：
影响因子：
7.4
通讯作者：
H. Kuriyama;H. Takano;L. Suzuki;H. Uchida;S. Kawano;H. Kuroiwa;T. Kuroiwa
中科院分区：
文献类型：
--
作者：
H. Kuriyama;H. Takano;L. Suzuki;H. Uchida;S. Kawano;H. Kuroiwa;T. Kuroiwa
仅在幼合子中表达的基因被认为在同花受精的绿色衣藻（Chlamyeliumreinhardtii）的发育中具有重要意义。代表Zys3基因的克隆从受精后10分钟使用受精卵制备的cDNA文库中分离。Zys3 cDNA克隆的测序导致两个相关的分子物种的分离。其中一个编码的蛋白质包含两种蛋白质间相互作用的基序，称为锚蛋白重复序列和WW结构域。另一个克隆缺乏锚蛋白重复序列，但在其他方面是相同的。受精后10 min，这些mRNA开始在细胞内同时积累，约4 h达到最高水平，此后水平显著下降。基因组DNA凝胶印迹分析表明Zys 3是一个单拷贝基因。Zys3蛋白在交配后2 h开始与Zys3 mRNA平行表达，在交配后6 h达到最高水平，但至少持续1 d。免疫细胞化学分析显示，它们的定位在内质网，这表明在内质网的形态变化或在合成和运输蛋白质的高尔基体或相关囊泡的作用。
Genes that are expressed only in the young zygote are considered to be of great importance in the development of an isogamous green alga, Chlamydomonas reinhardtii. Clones representing the Zys3 gene were isolated from a cDNA library prepared using zygotes at 10 min after fertilization. Sequencing of Zys3 cDNA clones resulted in the isolation of two related molecular species. One of them encoded a protein that contained two kinds of protein-to-protein interaction motifs known as ankyrin repeats and WW domains. The other clone lacked the ankyrin repeats but was otherwise identical. These mRNA species began to accumulate simultaneously in cells beginning 10 min after fertilization, and reached maximum levels at about 4 h, after which time levels decreased markedly. Genomic DNA gel-blot analysis indicated that Zys3 was a single-copy gene. The Zys3 proteins exhibited parallel expression to the Zys3 mRNAs at first, appearing 2 h after mating, and reached maximum levels at more than 6 h, but persisted to at least 1 d. Immunocytochemical analysis revealed their localization in the endoplasmic reticulum, which suggests a role in the morphological changes of the endoplasmic reticulum or in the synthesis and transport of proteins to the Golgi apparatus or related vesicles.