Characterization and function analysis of a novel gene, Hc-maoc-1, in the parasitic nematode Haemonochus contortus.
Characterization and function analysis of a novel gene, Hc-maoc-1, in the parasitic nematode Haemonochus contortus.
复制标题
寄生线虫捻转血矛线虫新基因 Hc-maoc-1 的表征和功能分析
DOI：
10.1186/s13071-017-1991-1
复制
发表时间：
2017-02-06
影响因子：
3.2
通讯作者：
Du A
中科院分区：
文献类型：
--
作者：
Ding H;Shi H;Shi Y;Guo X;Zheng X;Chen X;Zhou Q;Yang Y;Du A
在自由生活的秀丽隐杆线虫中，烯酰辅酶a水合酶（MAOC）是通过过氧化物酶体β-氧化来生物合成蛔虫苷脂肪酸衍生侧链所必需的。作为二脱氧糖的衍生物，ascarylose被用作daer信息素或daumones来诱导抗性daer幼虫阶段的发育。Hc-maoc-1基因是通过检索威康信托基金会桑格研究所的弯曲猴基因组数据库获得的。以不同发育阶段为模板，采用qRT-PCR方法分析Hc-maoc-1的转录水平。采用IFA法测定L3幼虫的表达规律，并用微注射法验证Hc-maoc-1 5′侧区启动子活性。采用过表达和RNAi实验对N2株进行了Hc-maoc-1基因功能的研究。Hc-maoc-1 cDNA全长900 bp，包含8个由7个内含子分隔的外显子，具有Hotdog结构域和maoc-样结构域，以及其他几个残基和一个水合酶2基序。该基因在整个生命周期中均有转录，并在弯纹夜蛾第4期幼虫（L4）达到转录高峰；而在滞育期L4，其转录水平降低。Hc-MAOC-1的蛋白表达和定位主要在L3幼虫的肠道中。在线虫中过表达Ce-maoc-1和Hc-maoc-1可以延长寿命和增加体型。Ce-MAOC-1和Hc-MAOC-1蛋白以点状模式定位于肠内。在秀丽隐杆线虫中，敲低ce - maac -1会缩短寿命和体长，减少幼崽数量，增加脂质储存。以秀丽隐杆线虫为模型生物，确定Hc-maoc-1在弯杆隐杆线虫中的功能。结果表明，与Ce-maoc-1具有相似的特性和功能，为Hc-maoc-1在黄花香薷daumones生物合成中的潜在功能提供了证据。本文的在线版本（doi:10.1186/s13071-017-1991-1）包含补充材料，可供授权用户使用。
Enoyl-CoA hydratase (MAOC) is required for the biosynthesis of the fatty acid-derive side chains of the ascaroside via peroxisome β-oxidation in the free-living nematode Caenorhabditis elegans. The derivative of dideoxy-sugar, ascarylose is used as dauer pheromones or daumones to induce development of the stress-resistant dauer larvae stage. Hc-maoc-1 gene was obtained by searching the Wellcome Trusts Sanger Institute’s H. contortus genomic database. qRT-PCR was performed to analyse the transcriptional levels of Hc-maoc-1 with different developmental stages as templates. IFA was carried out to determine the expression pattern in L3 larvae and micro-injection was used to verify the promoter activity of 5′-flanking region of Hc-maoc-1. Overexpression and RNAi experiments were applied in N2 strain to ascertain the gene function of Hc-maoc-1. The full-length cDNA of Hc-maoc-1 was 900 bp in length, which contained eight exons separated by seven introns and possessed the Hotdog domain and the MaoC-like domain, together with several other residues and a hydratase 2 motif. It was transcribed throughout the lifecycle and peaked in the fourth-stage larvae (L4) of H. contortus; however, its transcription level decreased in diapausing L4. The protein expression and location of Hc-MAOC-1 were mainly in the intestine of L3 larvae. Overexpression of Ce-maoc-1 and Hc-maoc-1 in C. elegans showed extended lifespan and increased body size. The protein Ce-MAOC-1 and Hc-MAOC-1 were localized in the intestine with a punctate pattern. In C. elegans, knockdown of Ce-maoc-1 conferred shortened lifespan and body lengths, decreased brood size and increased lipid storage. Caenorhabditis elegans was used as a model organism to ascertain the function of Hc-maoc-1 in H. contortus. Our results showed the similar characteristics and functions with Ce-maoc-1 and provided evidences of the potential functions of Hc-maoc-1 in biosynthesis of daumones in H. contortus. The online version of this article (doi:10.1186/s13071-017-1991-1) contains supplementary material, which is available to authorized users.