Innexin AGAP001476 Is Critical for Mediating Anti-Plasmodium Responses in Anopheles Mosquitoes

Li, Michelle W.M.; Wang, Jiuling; Zhao, Yang O.; Fikrig, Erol

doi:10.1074/jbc.m114.554519

Cited by 12 publications

(7 citation statements)

References 34 publications

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“…Gap junctions are present and active in nearly all insect physiological systems, where transfer of small molecules has been observed or is predicted to affect system processes including Malpighian tubule activity (Weng et al, 2008;Piermarini and Calkins, 2014), oocyte patterning and provisioning (Adler and Woodruff, 2000;Anderson and Woodruff, 2001;Bohrmann and Zimmermann, 2008;Kruger and Bohrmann, 2015) and electrical synapse connectivity (Phelan et al, 1996;Phelan et al, 2008;Anava et al, 2009;Anava et al, 2013). Gap junctions also have been observed in the immune system of insects, particularly during transient and continuous contact of haemocytes (Baerwald, 1975;Baerwald, 1979;Caveney and Berdan, 1982;Churchill et al, 1993), while functional data implicate them in haemolytic (Hasegawa et al, 2017) and gut immunity (Li et al, 2014b). At the same time, data indicate that hemichannels can form unapposed channels permitting direct exchange between the environment and cytoplasm (Luo and Turnbull, 2011), possibly playing a role in mediating apoptotic signals in haemocytes (Liu et al, 2013;Li et al, 2014a;Pang et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

Intracellular dynamics of polydnavirus innexin homologues

Hasegawa

Zhang

Turnbull

2020

Insect Molecular Biology

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Polydnaviruses associated with ichneumonid parasitoid wasps (Ichnoviruses) encode large numbers of genes, often in multigene families. The Ichnovirus Vinnexin gene family, which is expressed in parasitized lepidopteran larvae, encodes homologues of Innexins, the structural components of insect gap junctions. Here, we have examined intracellular behaviours of the Campoletis sonorensis Ichnovirus (CsIV) Vinnexins, alone and in combination with a host Innexin orthologue, Innexin2 (Inx2). QRT‐PCR verified that transcription of CsIV vinnexins occurs contemporaneously with inx2, implying co‐occurrence of Vinnexin and Inx2 proteins. Confocal microscopy demonstrated that epitope‐tagged VinnexinG (VnxG) and VinnexinQ2 (VnxQ2) exhibit similar subcellular localization as Spodoptera frugiperda Inx2 (Sf‐Inx2). Surface biotinylation assays verified that all three proteins localize to the cell surface, and cytochalasin B and nocodazole that they rely on actin and microtubule cytoskeletal networks for localization. Immunomicroscopy following co‐transfection of constructs indicates extensive co‐localization of Vinnexins with each other and Sf‐Inx2, and live‐cell imaging of mCherry‐labelled Inx2 supports that Vinnexins may affect Sf‐Inx2 distribution in a Vinnexin‐specific fashion. Our findings support that the Vinnexins may disrupt host cell physiology in a protein‐specific manner through altering gap junctional intercellular channel communication, as well as indirectly by affecting multicellular junction characteristics.

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Section: Discussionmentioning

confidence: 99%

Intracellular dynamics of polydnavirus innexin homologues

Hasegawa

Zhang

Turnbull

2020

Insect Molecular Biology

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“…Anopheles ribosomal protein S7 is routinely used as a reference gene in studies similar to this. There are also gene expression studies of Anopheles using actin ( Li et al, 2014 ; Luckhart et al, 1998 ) or EF1 ( Omondi, Majeed & Ignell, 2015 ) as a reference gene. However, most of these investigations utilized only a single reference gene for normalization, which is not recommended ( Bustin et al, 2009 ; Vandesompele et al, 2002 ).…”

Section: Discussionmentioning

confidence: 99%

Quantitative real-time PCR analysis of Anopheles dirus TEP1 and NOS during Plasmodium berghei infection, using three reference genes

Liew

Fong

Lau

2017

PeerJ

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Quantitative reverse transcription PCR (qRT-PCR) has been an integral part of characterizing the immunity of Anopheles mosquitoes towards Plasmodium invasion. Two anti-Plasmodium factors of Anopheles, thioester-containing protein 1 (TEP1) and nitric oxide synthase (NOS), play a role in the refractoriness of Anopheles towards Plasmodium infection and are generally expressed during infection. However, these are less studied in Anopheles dirus, a dominant malaria vector in Southeast Asia. Furthermore, most studies used a single reference gene for normalization during gene expression analysis without proper validation. This may lead to erroneous quantification of expression levels. Therefore, the present study characterized and investigated the expression profiles of TEP1 and NOS of Anopheles dirus during P. berghei infection. Prior to that, the elongation factor 1-alpha (EF1), actin 1 (Act) and ribosomal protein S7 (S7) genes were validated for their suitability as a set of reference genes. TEP1 and NOS expressions in An. dirus were found to be significantly induced after P. berghei infection.

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“…In the desert locust ( Schistocerca gregaria ), a decrease of expression levels in Inx1 (74%), Inx2 (85%), Inx3 (95%), and Inx4 (65%) genes was obtained compared to the controls [73]. In Anopheles mosquitoes, gene knockdown of innexin AGAP001476 mRNA gene was achieved with dsRNA at a 60% level [74].…”

Section: Inhibition Of Gap Junction Communicationmentioning

confidence: 99%

Gap Junction Channels of Innexins and Connexins: Relations and Computational Perspectives

Sánchez

Castro

Flores

et al. 2019

IJMS

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Gap junction (GJ) channels in invertebrates have been used to understand cell-to-cell communication in vertebrates. GJs are a common form of intercellular communication channels which connect the cytoplasm of adjacent cells. Dysregulation and structural alteration of the gap junction-mediated communication have been proven to be associated with a myriad of symptoms and tissue-specific pathologies. Animal models relying on the invertebrate nervous system have exposed a relationship between GJs and the formation of electrical synapses during embryogenesis and adulthood. The modulation of GJs as a therapeutic and clinical tool may eventually provide an alternative for treating tissue formation-related diseases and cell propagation. This review concerns the similarities between Hirudo medicinalis innexins and human connexins from nucleotide and protein sequence level perspectives. It also sets forth evidence of computational techniques applied to the study of proteins, sequences, and molecular dynamics. Furthermore, we propose machine learning techniques as a method that could be used to study protein structure, gap junction inhibition, metabolism, and drug development.

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Innexin AGAP001476 Is Critical for Mediating Anti-Plasmodium Responses in Anopheles Mosquitoes

Cited by 12 publications

References 34 publications

Intracellular dynamics of polydnavirus innexin homologues

Intracellular dynamics of polydnavirus innexin homologues

Quantitative real-time PCR analysis of Anopheles dirus TEP1 and NOS during Plasmodium berghei infection, using three reference genes

Gap Junction Channels of Innexins and Connexins: Relations and Computational Perspectives

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