Genetic screen for signal peptides in Hydra reveals novel secreted proteins and evidence for non-classical protein secretion
Genetic screen for signal peptides in Hydra reveals novel secreted proteins and evidence for non-classical protein secretion
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DOI：
10.1016/j.ejcb.2006.05.007
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发表时间：
2006-09-01
影响因子：
6.6
通讯作者：
David, Charles N.
中科院分区：
文献类型：
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作者：
Boettger, Angelika;Strasser, Dominik;David, Charles N.
我们已经使用酵母转化酶分泌载体pSUC筛选了Hydra cDNA文库中编码N-末端信号肽的序列[Jacobs等人，1997.用于分离编码分泌蛋白质的cDNA的遗传选择。Gene 198，289-296]。我们分离并测序了907个阳性克隆; 88%编码信号肽; 12%缺乏信号肽。通过搜索Hydra EST数据库，我们确定了所选克隆的全长序列。这些基因编码37个已知的信号肽蛋白和40个新的Hydra特异性信号肽蛋白。两个信号肽的序列，VEGF和铁蛋白，分泌途径的本地化被证实与GFP融合蛋白。此外，我们分离出105个克隆，缺乏信号肽，但支持转化酶分泌酵母。从这些克隆中分离质粒，并在转化酶阴性酵母细胞中再转化，证实了表型。一个这样的克隆编码足形态pedibin的GFP融合蛋白定位于转染水螅细胞的细胞质中，并没有进入ER/高尔基体分泌途径。pedibin和其他缺乏信号肽的蛋白质的分泌似乎通过非经典的蛋白质分泌途径发生。(c)2006年Elsevier GmbH。All rights reserved.
We have screened a Hydra cDNA library for sequences encoding N-terminal signal peptides using the yeast invertase secretion vector pSUC [Jacobs et al., 1997. A genetic selection for isolating cDNAs encoding secreted proteins. Gene 198, 289-296]. We isolated and sequenced 907 positive clones; 88% encoded signal peptides; 12% lacked signal peptides. By searching the Hydra EST database we identified full-length sequences for the selected clones. These encoded 37 known proteins with signal peptides and 40 novel Hydra-specific proteins with signal peptides. Localization of two signal peptide-containing sequences, VEGF and ferritin, to the secretory pathway was confirmed with GFP fusion proteins. In addition, we isolated 105 clones which lacked signal peptides but which supported invertase secretion from yeast. Isolation of plasmids from these clones and retransformation in invertase-negative yeast cells confirmed the phenotype. A GFP fusion protein of one such clone encoding the foot morphogen pedibin was localized to the cytoplasm in transfected Hydra cells and did not enter the ER/Golgi secretory pathway. Secretion of pedibin and other proteins lacking signal peptides appears to occur by a non-classical protein secretion route. (c) 2006 Elsevier GmbH. All rights reserved.