Abstract
BackgroundEnoyl-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.MethodsHc-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.ResultsThe 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.ConclusionCaenorhabditis 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.
Highlights
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
Characterization of cDNA and phylogenetic analysis of amino acid sequence The full length of Hc-maoc-1 was 1,777 bp and contained 8 exons separated by 7 introns (Fig. 1)
The phylogenetic tree showed that HcMAOC-1 was in the same clad with C. elegans, which indicated a highly similarity between Hc-Enoyl-CoA hydrate (MAOC)-1 and Ce-MAOC-1 in amino acids
Summary
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. The gastric nematode Haemonchus contortus infects small ruminants (sheep and goats) worldwide, and causes great production losses. Infection by the infective third stage larvae (iL3) is seasonal. A form of arrested development, occurs in the early fourth stage of H. contortus in abomasa of ruminants [2]. It is a strategy for this parasitic nematode to adapt to adverse environmental conditions such as low temperature in winter, low oxygen or immunoreaction of the infected host [1,2,3]
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