Functional Identification of Complement Factor D and Analysis of Its Expression during GCRV Infection in Grass Carp (<i>Ctenopharyngodon</i> <i>idella</i>)

Complement factor D (Df) is a serine protease well known for activating the alternative pathway (AP) in mammals by promoting the cleavage of complement component 3 (C3), thus becoming involved in innate defense. In teleost fish, however, the functional mechanisms of Df in the AP and against pathogen...

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Autores principales:	Chunhua Ding, Tiaoyi Xiao, Beibei Qin, Baohong Xu, Zhao Lv, Hongquan Wang
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	complement factor D <i>Ctenopharyngodon idella</i> cleavage of C3 GCRV expression patterns Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/8325d381c9dd45eeaf7dc6efa60e147d
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spelling	oai:doaj.org-article:8325d381c9dd45eeaf7dc6efa60e147d2021-11-11T17:25:40ZFunctional Identification of Complement Factor D and Analysis of Its Expression during GCRV Infection in Grass Carp (<i>Ctenopharyngodon</i> <i>idella</i>)10.3390/ijms2221120111422-00671661-6596https://doaj.org/article/8325d381c9dd45eeaf7dc6efa60e147d2021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/12011https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Complement factor D (Df) is a serine protease well known for activating the alternative pathway (AP) in mammals by promoting the cleavage of complement component 3 (C3), thus becoming involved in innate defense. In teleost fish, however, the functional mechanisms of Df in the AP and against pathogen infection are far from clear. In the present study, we cloned and characterized the Df gene, <i>Ci</i>Df, from grass carp (<i>Ctenopharyngodon idella</i>) and analyzed its function in promoting C3 cleavage and expression changes after grass carp reovirus (GCRV) infection. The open reading frame of <i>Ci</i>Df was found to be 753 bp, encoding 250 amino acids with a molecular mass of 27.06 kDa. <i>Ci</i>Df harbors a conserved Tryp_SPc domain, with three conserved residues representing the catalytic triad and three conserved binding sites in the substrate specificity pocket. Pairwise alignment showed that <i>Ci</i>Df shares the highest identity (96%) and similarity (98%) with Df from <i>Anabarilius grahami</i>. Phylogenetic analysis indicated that <i>Ci</i>Df and other fish Dfs formed a distinct evolutionary branch. Similar to most Dfs from other vertebrates, the <i>Ci</i>Df gene structure is characterized by four introns and five exons. The incubation of recombinant <i>Ci</i>Df protein with grass carp serum significantly increased the C3b content, demonstrating the conserved function of <i>Ci</i>Df in the AP in promoting C3 cleavage, similar to Dfs in mammals. <i>Ci</i>Df mRNA expression was widely detected in various tissues and levels were relatively higher in the liver, spleen, and intestine of grass carp. During GCRV infection over a 168-hour period, a high level of <i>Ci</i>Df mRNA expression in the liver, spleen, and intestine was maintained at 144 and 168 h, suggesting AP activity at the late stage of GCRV infection. Collectively, the above results reveal the conserved structure and function of <i>Ci</i>Df and its distinct expression patterns after GCRV infection, which provide a key basis for studying the roles of Df and AP during GCRV infection in the grass carp <i>C. idella</i>.Chunhua DingTiaoyi XiaoBeibei QinBaohong XuZhao LvHongquan WangMDPI AGarticlecomplement factor D<i>Ctenopharyngodon idella</i>cleavage of C3GCRVexpression patternsBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 12011, p 12011 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	complement factor D <i>Ctenopharyngodon idella</i> cleavage of C3 GCRV expression patterns Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	complement factor D <i>Ctenopharyngodon idella</i> cleavage of C3 GCRV expression patterns Biology (General) QH301-705.5 Chemistry QD1-999 Chunhua Ding Tiaoyi Xiao Beibei Qin Baohong Xu Zhao Lv Hongquan Wang Functional Identification of Complement Factor D and Analysis of Its Expression during GCRV Infection in Grass Carp (<i>Ctenopharyngodon</i> <i>idella</i>)
description	Complement factor D (Df) is a serine protease well known for activating the alternative pathway (AP) in mammals by promoting the cleavage of complement component 3 (C3), thus becoming involved in innate defense. In teleost fish, however, the functional mechanisms of Df in the AP and against pathogen infection are far from clear. In the present study, we cloned and characterized the Df gene, <i>Ci</i>Df, from grass carp (<i>Ctenopharyngodon idella</i>) and analyzed its function in promoting C3 cleavage and expression changes after grass carp reovirus (GCRV) infection. The open reading frame of <i>Ci</i>Df was found to be 753 bp, encoding 250 amino acids with a molecular mass of 27.06 kDa. <i>Ci</i>Df harbors a conserved Tryp_SPc domain, with three conserved residues representing the catalytic triad and three conserved binding sites in the substrate specificity pocket. Pairwise alignment showed that <i>Ci</i>Df shares the highest identity (96%) and similarity (98%) with Df from <i>Anabarilius grahami</i>. Phylogenetic analysis indicated that <i>Ci</i>Df and other fish Dfs formed a distinct evolutionary branch. Similar to most Dfs from other vertebrates, the <i>Ci</i>Df gene structure is characterized by four introns and five exons. The incubation of recombinant <i>Ci</i>Df protein with grass carp serum significantly increased the C3b content, demonstrating the conserved function of <i>Ci</i>Df in the AP in promoting C3 cleavage, similar to Dfs in mammals. <i>Ci</i>Df mRNA expression was widely detected in various tissues and levels were relatively higher in the liver, spleen, and intestine of grass carp. During GCRV infection over a 168-hour period, a high level of <i>Ci</i>Df mRNA expression in the liver, spleen, and intestine was maintained at 144 and 168 h, suggesting AP activity at the late stage of GCRV infection. Collectively, the above results reveal the conserved structure and function of <i>Ci</i>Df and its distinct expression patterns after GCRV infection, which provide a key basis for studying the roles of Df and AP during GCRV infection in the grass carp <i>C. idella</i>.
format	article
author	Chunhua Ding Tiaoyi Xiao Beibei Qin Baohong Xu Zhao Lv Hongquan Wang
author_facet	Chunhua Ding Tiaoyi Xiao Beibei Qin Baohong Xu Zhao Lv Hongquan Wang
author_sort	Chunhua Ding
title	Functional Identification of Complement Factor D and Analysis of Its Expression during GCRV Infection in Grass Carp (<i>Ctenopharyngodon</i> <i>idella</i>)
title_short	Functional Identification of Complement Factor D and Analysis of Its Expression during GCRV Infection in Grass Carp (<i>Ctenopharyngodon</i> <i>idella</i>)
title_full	Functional Identification of Complement Factor D and Analysis of Its Expression during GCRV Infection in Grass Carp (<i>Ctenopharyngodon</i> <i>idella</i>)
title_fullStr	Functional Identification of Complement Factor D and Analysis of Its Expression during GCRV Infection in Grass Carp (<i>Ctenopharyngodon</i> <i>idella</i>)
title_full_unstemmed	Functional Identification of Complement Factor D and Analysis of Its Expression during GCRV Infection in Grass Carp (<i>Ctenopharyngodon</i> <i>idella</i>)
title_sort	functional identification of complement factor d and analysis of its expression during gcrv infection in grass carp (<i>ctenopharyngodon</i> <i>idella</i>)
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/8325d381c9dd45eeaf7dc6efa60e147d
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Functional Identification of Complement Factor D and Analysis of Its Expression during GCRV Infection in Grass Carp (<i>Ctenopharyngodon</i> <i>idella</i>)

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