Functional characterization of two clip domain serine proteases in innate immune responses of Aedes aegypti

Abstract Background Clip domain serine proteases (CLIPs), a very diverse group of proteolytic enzymes, play a crucial role in the innate immunity of insects. Innate immune responses are the first line of defense in mosquitoes against the invasion of pathogenic microorganisms. The Toll pathway, immun...

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Autores principales:	Hao-Cheng Wang, Qiu-Hui Wang, Biswajit Bhowmick, Yi-Xun Li, Qian Han
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Innate immunity Aedes aegypti Prophenoloxidase RNA interference Clip domain serine proteases Microbial infection Infectious and parasitic diseases RC109-216
Acceso en línea:	https://doaj.org/article/e861f222d8c040d386d65efb80f6bf70
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spelling	oai:doaj.org-article:e861f222d8c040d386d65efb80f6bf702021-11-28T12:22:54ZFunctional characterization of two clip domain serine proteases in innate immune responses of Aedes aegypti10.1186/s13071-021-05091-91756-3305https://doaj.org/article/e861f222d8c040d386d65efb80f6bf702021-11-01T00:00:00Zhttps://doi.org/10.1186/s13071-021-05091-9https://doaj.org/toc/1756-3305Abstract Background Clip domain serine proteases (CLIPs), a very diverse group of proteolytic enzymes, play a crucial role in the innate immunity of insects. Innate immune responses are the first line of defense in mosquitoes against the invasion of pathogenic microorganisms. The Toll pathway, immunodeficiency (IMD) pathway and melanization are the main processes of innate immunity in Aedes aegypti. CLIPS are classified into five subfamilies—CLIPA, CLIPB, CLIPC, CLIPD, and CLIPE—based on their sequence specificity and phylogenetic relationships. We report the functional characterization of the genes that code for two CLIPs in Ae. aegypti (Ae): Ae-CLIPB15 and Ae-CLIPB22. Methods Clustal Omega was used for multiple amino acid sequence alignment of Ae-CLIPB15 and Ae-CLIPB22 with different CLIP genes from other insect species. The spatiotemporal expression profiles of Ae-CLIPB15 and Ae-CLIPB22 were examined. We determined whether Ae-CLIPB15 and Ae-CLIPB22 respond to microbial challenge and tissue injury. RNA interference (RNAi) was used to explore the function of Ae-CLIPB15 and Ae-CLIPB22 in the defense of Ae. aegypti against bacterial and fungal infections. The expression levels of nuclear factor kappa B (NF-κB) transcription factors REL1 and REL2 in the Toll pathway and IMD pathway after bacterial infection were investigated. Finally, the change in phenoloxidase (PO) activity in Ae-CLIPB15 and Ae-CLIPB22 knockdown adults was investigated. Results We performed spatiotemporal gene expression profiling of Ae-CLIPB15 and Ae-CLIPB22 genes in Ae. aegypti using quantitative real-time polymerase chain reaction. These genes were expressed in different stages and tissues. The messenger RNA (mRNA) levels for both genes were also up-regulated by Gram-negative bacteria Escherichia coli, Gram-positive bacteria Staphylococcus aureus and fungal Beauveria bassiana infections, as well as in the tissue injury experiments. RNAi-mediated knockdown of Ae-CLIPB15 led to a significant decrease of PO activity in the hemolymph of Ae. aegypti, while other RNAi experiments revealed that both Ae-CLIPB15 and Ae-CLIPB22 were involved in immune defense against bacterial and fungal infections. The mRNA expression of NF-κB transcription factors REL1 and REL2 in the Toll pathway and IMD pathway differed between Ae-CLIPB15 and Ae-CLIPB22 knockdown mosquitoes infected with bacteria and wild type mosquitoes infected with bacteria. Conclusions Our findings suggest that Ae-CLIPB15 and Ae-CLIPB22 play a critical role in mosquito innate immunity, and that they are involved in immune responses to injury and infection. Their regulation of transcription factors and PO activity indicates that they also play a specific role in the regulation of innate immunity. Graphical AbstractHao-Cheng WangQiu-Hui WangBiswajit BhowmickYi-Xun LiQian HanBMCarticleInnate immunityAedes aegyptiProphenoloxidaseRNA interferenceClip domain serine proteasesMicrobial infectionInfectious and parasitic diseasesRC109-216ENParasites & Vectors, Vol 14, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Innate immunity Aedes aegypti Prophenoloxidase RNA interference Clip domain serine proteases Microbial infection Infectious and parasitic diseases RC109-216
spellingShingle	Innate immunity Aedes aegypti Prophenoloxidase RNA interference Clip domain serine proteases Microbial infection Infectious and parasitic diseases RC109-216 Hao-Cheng Wang Qiu-Hui Wang Biswajit Bhowmick Yi-Xun Li Qian Han Functional characterization of two clip domain serine proteases in innate immune responses of Aedes aegypti
description	Abstract Background Clip domain serine proteases (CLIPs), a very diverse group of proteolytic enzymes, play a crucial role in the innate immunity of insects. Innate immune responses are the first line of defense in mosquitoes against the invasion of pathogenic microorganisms. The Toll pathway, immunodeficiency (IMD) pathway and melanization are the main processes of innate immunity in Aedes aegypti. CLIPS are classified into five subfamilies—CLIPA, CLIPB, CLIPC, CLIPD, and CLIPE—based on their sequence specificity and phylogenetic relationships. We report the functional characterization of the genes that code for two CLIPs in Ae. aegypti (Ae): Ae-CLIPB15 and Ae-CLIPB22. Methods Clustal Omega was used for multiple amino acid sequence alignment of Ae-CLIPB15 and Ae-CLIPB22 with different CLIP genes from other insect species. The spatiotemporal expression profiles of Ae-CLIPB15 and Ae-CLIPB22 were examined. We determined whether Ae-CLIPB15 and Ae-CLIPB22 respond to microbial challenge and tissue injury. RNA interference (RNAi) was used to explore the function of Ae-CLIPB15 and Ae-CLIPB22 in the defense of Ae. aegypti against bacterial and fungal infections. The expression levels of nuclear factor kappa B (NF-κB) transcription factors REL1 and REL2 in the Toll pathway and IMD pathway after bacterial infection were investigated. Finally, the change in phenoloxidase (PO) activity in Ae-CLIPB15 and Ae-CLIPB22 knockdown adults was investigated. Results We performed spatiotemporal gene expression profiling of Ae-CLIPB15 and Ae-CLIPB22 genes in Ae. aegypti using quantitative real-time polymerase chain reaction. These genes were expressed in different stages and tissues. The messenger RNA (mRNA) levels for both genes were also up-regulated by Gram-negative bacteria Escherichia coli, Gram-positive bacteria Staphylococcus aureus and fungal Beauveria bassiana infections, as well as in the tissue injury experiments. RNAi-mediated knockdown of Ae-CLIPB15 led to a significant decrease of PO activity in the hemolymph of Ae. aegypti, while other RNAi experiments revealed that both Ae-CLIPB15 and Ae-CLIPB22 were involved in immune defense against bacterial and fungal infections. The mRNA expression of NF-κB transcription factors REL1 and REL2 in the Toll pathway and IMD pathway differed between Ae-CLIPB15 and Ae-CLIPB22 knockdown mosquitoes infected with bacteria and wild type mosquitoes infected with bacteria. Conclusions Our findings suggest that Ae-CLIPB15 and Ae-CLIPB22 play a critical role in mosquito innate immunity, and that they are involved in immune responses to injury and infection. Their regulation of transcription factors and PO activity indicates that they also play a specific role in the regulation of innate immunity. Graphical Abstract
format	article
author	Hao-Cheng Wang Qiu-Hui Wang Biswajit Bhowmick Yi-Xun Li Qian Han
author_facet	Hao-Cheng Wang Qiu-Hui Wang Biswajit Bhowmick Yi-Xun Li Qian Han
author_sort	Hao-Cheng Wang
title	Functional characterization of two clip domain serine proteases in innate immune responses of Aedes aegypti
title_short	Functional characterization of two clip domain serine proteases in innate immune responses of Aedes aegypti
title_full	Functional characterization of two clip domain serine proteases in innate immune responses of Aedes aegypti
title_fullStr	Functional characterization of two clip domain serine proteases in innate immune responses of Aedes aegypti
title_full_unstemmed	Functional characterization of two clip domain serine proteases in innate immune responses of Aedes aegypti
title_sort	functional characterization of two clip domain serine proteases in innate immune responses of aedes aegypti
publisher	BMC
publishDate	2021
url	https://doaj.org/article/e861f222d8c040d386d65efb80f6bf70
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Functional characterization of two clip domain serine proteases in innate immune responses of Aedes aegypti

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