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Characterization and function analysis of a novel gene, Hc-maoc-1, in the parasitic nematode Haemonochus contortus
BACKGROUND: 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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5294872/ https://www.ncbi.nlm.nih.gov/pubmed/28166831 http://dx.doi.org/10.1186/s13071-017-1991-1 |
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author | Ding, Haojie Shi, Hengzhi Shi, Yu Guo, Xiaolu Zheng, Xiuping Chen, Xueqiu Zhou, Qianjin Yang, Yi Du, Aifang |
author_facet | Ding, Haojie Shi, Hengzhi Shi, Yu Guo, Xiaolu Zheng, Xiuping Chen, Xueqiu Zhou, Qianjin Yang, Yi Du, Aifang |
author_sort | Ding, Haojie |
collection | PubMed |
description | BACKGROUND: 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. METHODS: 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 N(2) strain to ascertain the gene function of Hc-maoc-1. RESULTS: 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. CONCLUSION: 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13071-017-1991-1) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5294872 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-52948722017-02-09 Characterization and function analysis of a novel gene, Hc-maoc-1, in the parasitic nematode Haemonochus contortus Ding, Haojie Shi, Hengzhi Shi, Yu Guo, Xiaolu Zheng, Xiuping Chen, Xueqiu Zhou, Qianjin Yang, Yi Du, Aifang Parasit Vectors Research BACKGROUND: 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. METHODS: 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 N(2) strain to ascertain the gene function of Hc-maoc-1. RESULTS: 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. CONCLUSION: 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13071-017-1991-1) contains supplementary material, which is available to authorized users. BioMed Central 2017-02-06 /pmc/articles/PMC5294872/ /pubmed/28166831 http://dx.doi.org/10.1186/s13071-017-1991-1 Text en © The Author(s). 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Ding, Haojie Shi, Hengzhi Shi, Yu Guo, Xiaolu Zheng, Xiuping Chen, Xueqiu Zhou, Qianjin Yang, Yi Du, Aifang Characterization and function analysis of a novel gene, Hc-maoc-1, in the parasitic nematode Haemonochus contortus |
title | Characterization and function analysis of a novel gene, Hc-maoc-1, in the parasitic nematode Haemonochus contortus |
title_full | Characterization and function analysis of a novel gene, Hc-maoc-1, in the parasitic nematode Haemonochus contortus |
title_fullStr | Characterization and function analysis of a novel gene, Hc-maoc-1, in the parasitic nematode Haemonochus contortus |
title_full_unstemmed | Characterization and function analysis of a novel gene, Hc-maoc-1, in the parasitic nematode Haemonochus contortus |
title_short | Characterization and function analysis of a novel gene, Hc-maoc-1, in the parasitic nematode Haemonochus contortus |
title_sort | characterization and function analysis of a novel gene, hc-maoc-1, in the parasitic nematode haemonochus contortus |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5294872/ https://www.ncbi.nlm.nih.gov/pubmed/28166831 http://dx.doi.org/10.1186/s13071-017-1991-1 |
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