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A signal peptide secretion screen in Fucus distichus embryos reveals expression of glucanase, EGF domain-containing, and LRR receptor kinase-like polypeptides during asymmetric cell growth

机译:胚胎中的信号肽分泌筛分揭示了在不对称细胞生长期间葡聚糖酶,含EGF结构域和LRR受体激酶样多肽的表达

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摘要

Zygotes of the brown alga Fucus distichus (L.) Powell develop polarity prior to the first embryonic cell division and retain a pattern of asymmetric growth during early embryogenesis. In order to identify F. distichus polypeptides secreted during asymmetric cell growth, we used a functional assay in Saccharomyces cerevisiae to screen a cDNA library generated from asymmetrically growing Fucus embryos for sequences encoding polypeptides that function as signal peptides for secretion. We isolated and sequenced 222 plasmids containing Fucus cDNAs encoding signal peptide activity. The cDNA inserts from these plasmids were translated in silico into 244 potential polypeptide sequences, 169 of which are predicted to contain signal peptides. BlastP analysis of the Fucus sequences revealed similarity between many Fucus proteins and cell surface proteins that function in development in other eukaryotes, including epidermal growth factor (EGF)-like repeat-containing proteins, plant leucine-rich repeat (LRR)-receptor kinases, and algal β-1, 3-exoglucanase. However, most of the isolated Fucus polypeptides lack similarity to known proteins. The isolation of cDNAs encoding secreted Fucus proteins provides an important step toward characterizing cell surface proteins important for asymmetric organization and growth in fucoid embryos.
机译:棕榈藻藻曲板的Zygotes在第一胚胎细胞分裂之前发育极性,并在早期胚胎发生期间保留不对称生长的图案。为了鉴定在不对称细胞生长期间分泌的DisiChus多肽,我们在酿酒酵母中使用了功能测定,以筛选由不对称生长的黄糖胚胎产生的cDNA文库,用于编码作为分泌的信号肽的多肽。我们被隔离和测序含有发病率CDNA的222个质粒,编码信号肽活性。将来自这些质粒的cDNA插入物在硅中翻译成244个潜在的多肽序列,其中169例预计含有信号肽。发病气囊序列的BLASTP分析显示了许多心脏蛋白和细胞表面蛋白之间的相似性,其在其他真核生物中发挥作用,包括表皮生长因子(EGF) - 含有含重复的蛋白质,植物富含富含植物的重复(LRR) - 预应体激酶,和藻类β-1,3-外葡聚糖酶。然而,大多数分离的阴影多肽缺乏与已知蛋白质相似之处。编码分泌的心脏蛋白的CDNA的分离为表征对不对称组织和生长在粘合剂胚胎中的细胞表面蛋白质的重要​​步骤提供了重要的步骤。

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