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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...

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Autores principales: Ding, Haojie, Shi, Hengzhi, Shi, Yu, Guo, Xiaolu, Zheng, Xiuping, Chen, Xueqiu, Zhou, Qianjin, Yang, Yi, Du, Aifang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
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.
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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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