Phenotype-based clustering of glycosylation-related genes by RNAi-mediated gene silencing.
Phenotype-based clustering of glycosylation-related genes by RNAi-mediated gene silencing.
复制标题
DOI：
10.1111/gtc.12246
复制
发表时间：
2015-06
期刊：
影响因子：
--
通讯作者：
Goto S
中科院分区：
文献类型：
--
作者：
Yamamoto-Hino M;Yoshida H;Ichimiya T;Sakamura S;Maeda M;Kimura Y;Sasaki N;Aoki-Kinoshita KF;Kinoshita-Toyoda A;Toyoda H;Ueda R;Nishihara S;Goto S
聚糖结构通过糖基化相关（GR）蛋白（如糖基转移酶、聚糖修饰酶和核苷酸-糖转运蛋白）进行的一系列反应合成。例如，糖胺聚糖（GAG）的共同核心区由肽-O-木糖基转移酶、β 1，4-半乳糖基转移酶I、β 1，3-半乳糖基转移酶II和β 1，3-葡糖醛酸基转移酶顺序合成。这提出了参与相同聚糖合成的GR蛋白的功能损伤可能导致相同表型异常的可能性。为了研究这种可能性，在果蝇中进行了GR和蛋白聚糖核心蛋白编码基因的全面沉默。果蝇GR候选基因（125个）被分为5个功能组，用于合成GAG，N-连接，O-连接，Notch相关和未知聚糖。时空调节沉默引起了一系列畸形的表型，分为三种类型：额外的静脉，厚静脉，脱色。聚类表型反映了GAG、Notch上的Fringe依赖性聚糖和位于非还原末端或附近的聚糖（本文称为聚糖的末端结构域）的生物合成途径。基于表型聚类，预测CG 33145参与末端结构域的形成。我们进一步的分析表明，CG 33145在末端N-连接聚糖的合成中表现出半乳糖基转移酶活性。因此，表型聚类具有潜在的新GR基因的功能预测。
Glycan structures are synthesized by a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters. For example, the common core region of glycosaminoglycans (GAGs) is sequentially synthesized by peptide-O-xylosyltransferase, β1,4-galactosyltransferase I, β1,3-galactosyltransferase II, and β1,3-glucuronyltransferase. This raises the possibility that functional impairment of GR proteins involved in synthesis of the same glycan might result in the same phenotypic abnormality. To examine this possibility, comprehensive silencing of genes encoding GR and proteoglycan core proteins was conducted in Drosophila. Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. Spatiotemporally regulated silencing caused a range of malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation. The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). Based on the phenotypic clustering, CG33145 was predicted to be involved in formation of terminal domains. Our further analysis showed that CG33145 exhibited galactosyltransferase activity in synthesis of terminal N-linked glycans. Phenotypic clustering, therefore, has potential for the functional prediction of novel GR genes.