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首页> 外文期刊>Harmful Algae >Transcriptome sequencing reveals single domain Type I-like polyketide synthases in the toxic dinoflagellate Gambierdiscus polynesiensis
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Transcriptome sequencing reveals single domain Type I-like polyketide synthases in the toxic dinoflagellate Gambierdiscus polynesiensis

机译：转录组测序揭示了有毒的鞭毛鞭毛冈比亚鞭毛虫中的单结构域I型聚酮化合物合酶。

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摘要

Ciguatoxins (CTXs) are potent neurotoxins responsible for the food-borne illness known as ciguatera that occurs after consumption of contaminated fish. Benthic dinoflagellates of the genus Gambierdiscus spp. are known as the main producers of CTXs. CTXs are polycyclic polyethers, presumed to be synthesized by polyketide synthase (PKS) complexes; however, the mechanisms of CTX biosynthesis remain unresolved. Here, we investigated a de novo transcriptome assembly of Gambierdiscus polynesiensis TB-92 clone, a highly toxic producer of Pacific ciguatoxins, and focused on the identification of PKS transcripts. A cDNA library generated using a spliced leader (SL) priming approach, which specifically targets the dinoflagellate nuclear transcriptome, was sequenced by Roche 454. This strategy produced 1,221,335 raw reads, assembled into 16,336 unique contigs. Contigs were subjected to BLAST search, annotated with Gene Ontology (GO) terms and enriched with enzyme codes (EC) from Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Thirty-three PKS-related sequences were thus identified. Twenty-two contigs encoded single domain β-ketoacyl synthases (KS) with sequence similarity to Type Ⅰ PKSs, as reported in other dinoflagellates. A conserved motif previously observed near the 5' end of dinoflagellate KS domain transcripts was present in G. polynesiensis, and distinguished two groups of KS domain sequences. Ketoreductase (KR), acyltransferase (AT), and acyl carrier protein (ACP) domains were also found on single domain containing transcripts. KEGG pathway mapping placed three of the KS sequences containing the PKS conserved domain (cd00833) in the fatty acid biosynthesis pathway. No contigs were found encoding the conserved domains typically found in elongating ketosynthase domains of fatty acid synthases (cd00832, Type Ⅰ or cd00834, Type Ⅱ). Contigs mapping to other parts of the fatty acid biosynthesis pathway similarly encoded individual domains, suggesting that fatty acid synthesis takes place in multiprotein complexes. Other than the three KS domains, none of the sequences mapping to the fatty acid biosynthesis pathway overlapped with those annotated as PKSs. These data lend support to the idea that PKSs may contribute to both polyketide and fatty acid synthesis in dinoflagellates. This dataset provides important background to future research in order to understand the complex mechanism of toxin production in this dinoflagellate.
机译：Ciguatoxins（CTX）是强效神经毒素，可导致食用被污染鱼后发生的食源性疾病，称为Ciguatera。 Gambierdiscus spp属的底栖鞭毛虫。被称为CTX的主要生产商。 CTX是多环聚醚，推测是由聚酮化合物合酶（PKS）配合物合成的；然而，CTX生物合成的机制仍未解决。在这里，我们研究了太平洋雪茄毒素的高毒性生产者，Gambierdiscus polynesiensis TB-92克隆的从头转录组装配，并着重于PKS转录本的鉴定。罗氏454对使用特异靶向鞭毛鞭毛核转录组的剪接前导（SL）引物方法生成的cDNA文库进行了测序。该策略产生了1,221,335个原始读段，组装成16,336个独特的重叠群。对重叠群进行了BLAST搜索，并注有基因本体论（GO）术语，并富含《京都议定书》的基因和基因组百科全书（KEGG）数据库中的酶代码（EC）。因此鉴定了33个与PKS相关的序列。 22个重叠群编码的单结构域β-酮酰基合酶（KS）与Ⅰ型PKSs具有序列相似性，如其他鞭毛虫中报道的那样。以前在聚鞭毛的KS域转录物的5'端附近观察到的保守基序存在于波利尼西亚草中，并区分了两组KS域序列。酮还原酶（KR），酰基转移酶（AT）和酰基载体蛋白（ACP）结构域也在包含单个结构域的转录本上发现。 KEGG途径作图法将三个包含PKS保守域（cd00833）的KS序列中的三个置于脂肪酸生物合成途径中。没有发现重叠群，其编码通常在脂肪酸合酶的延伸酮合成酶结构域（cd00832，Ⅰ型或cd00834，Ⅱ型）中发现的保守结构域。重叠群映射到脂肪酸生物合成途径的其他部分类似地编码单个结构域，表明脂肪酸合成发生在多蛋白复合物中。除三个KS结构域外，映射至脂肪酸生物合成途径的序列均未与注释为PKS的序列重叠。这些数据支持了PKSs可能同时促进鞭毛藻中聚酮化合物和脂肪酸合成的想法。该数据集为将来的研究提供了重要背景，以了解该鞭毛甲藻毒素产生的复杂机制。

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  • 来源
    《Harmful Algae》 |2014年第6期|29-37|共9页
  • 作者

    R. Pawlowiez; J.S. Morey; H.T. Darius; M. Chinain; F.M. Van Dolah;

  • 作者单位

    UMR241-EIO, Laboratoire des Microalgues Toxiques, Institut Louis Malarde, BP30, Papeete-Tahiti 98713, French Polynesia;

    Marine Biotoxins Program, NOAA, Center for Coastal and Environmental Health and Biomolecular Research, 219 Fort Johnson Road, Charleston, SC 29412, USA;

    UMR241-EIO, Laboratoire des Microalgues Toxiques, Institut Louis Malarde, BP30, Papeete-Tahiti 98713, French Polynesia;

    UMR241-EIO, Laboratoire des Microalgues Toxiques, Institut Louis Malarde, BP30, Papeete-Tahiti 98713, French Polynesia;

    Marine Biotoxins Program, NOAA, Center for Coastal and Environmental Health and Biomolecular Research, 219 Fort Johnson Road, Charleston, SC 29412, USA;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Gambierdiscus polynesiensis; Dinoflagellate; Toxin biosynthesis; Transcriptomic analysis; Polyketide synthase; Fatty acid;

    机译：Gambierdiscus polynesiensis;鞭毛藻;毒素生物合成;转录组分析;聚酮化合物合酶;脂肪酸;

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