Genetic evidence of a redox-dependent systemic wound response via Hayan Protease-Phenoloxidase system inDrosophila

Nam, Hyuck‐Jin; Jang, In-Hwan; You, Hyejin; Lee, Kyung‐Ah; Lee, Won–Jae

doi:10.1038/emboj.2011.476

Cited by 106 publications

(141 citation statements)

References 60 publications

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“…This blackening reaction results from de novo synthesis of melanin by PO and is also enhanced by the presence of microbial products [13]. We investigated the role of PPO1 and PPO2 in this process by observing the melanization site 30 min after clean injury in larvae ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Prophenoloxidase Activation Is Required for Survival to Microbial Infections in Drosophila

et al. 2014

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The melanization reaction is a major immune response in Arthropods and involves the rapid synthesis of melanin at the site of infection and injury. A key enzyme in the melanization process is phenoloxidase (PO), which catalyzes the oxidation of phenols to quinones, which subsequently polymerize into melanin. The Drosophila genome encodes three POs, which are primarily produced as zymogens or prophenoloxidases (PPO). Two of them, PPO1 and PPO2, are produced by crystal cells. Here we have generated flies carrying deletions in PPO1 and PPO2. By analyzing these mutations alone and in combination, we clarify the functions of both PPOs in humoral melanization. Our study shows that PPO1 and PPO2 are responsible for all the PO activity in the hemolymph. While PPO1 is involved in the rapid early delivery of PO activity, PPO2 is accumulated in the crystals of crystal cells and provides a storage form that can be deployed in a later phase. Our study also reveals an important role for PPO1 and PPO2 in the survival to infection with Gram-positive bacteria and fungi, underlining the importance of melanization in insect host defense.

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Section: Resultsmentioning

confidence: 99%

Prophenoloxidase Activation Is Required for Survival to Microbial Infections in Drosophila

et al. 2014

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“…Future work will enable us to localize the differential regulation of genes/pathways to individual cells/tissues. The site of damage, neighboring tissues [46] and more distant sites [47,48] are potential targets. Our functional analysis shows that IDGF3 supports a swift recovery after wounding.…”

Section: Discussionmentioning

confidence: 99%

The Drosophila Chitinase-Like Protein IDGF3 Is Involved in Protection against Nematodes and in Wound Healing

et al. 2015

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required for the formation of hemolymph clots that seal wounds, and Idgf3 mutants display an extended developmental delay during wound healing. Altogether, our findings indicate that vertebrate and invertebrate CLP proteins function in analogous settings and have a broad impact on inflammatory reactions and infections. This opens the way to further genetic analysis of Drosophila IDGF3 and will help to elucidate the exact molecular context of CLP function.

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“…As a complement to the PO assay, a Western Blot using antibodies against Drosophila PO1 and PO2 can be performed on larval or adult hemolymph samples [97,131]. By this method, both the naïve PPO (75 kDa) and the mature form of PO (70 kDa), resulting from PO activation upon proteolytic cleavage by the upstream serine protease Hayan, can be detected.…”

Section: Ppo Cleavage Assaymentioning

confidence: 99%

Methods to study Drosophila immunity

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Bretscher

Binggeli

et al. 2014

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a b s t r a c tInnate immune mechanisms are well conserved throughout evolution, and many theoretical concepts, molecular pathways and gene networks are applicable to invertebrate model organisms as much as vertebrate ones. Drosophila immunity research benefits from an easily manipulated genome, a fantastic international resource of transgenic tools and over a quarter century of accumulated techniques and approaches to study innate immunity. Here we present a short collection of ways to challenge the fruit fly immune system with various pathogens and parasites, as well as read-outs to assess its functions, including cellular and humoral immune responses. Our review covers techniques for assessing the kinetics and efficiency of immune responses quantitatively and qualitatively, such as survival analysis, bacterial persistence, antimicrobial peptide gene expression, phagocytosis and melanisation assays. Finally, we offer a toolkit of Drosophila strains available to the research community for current and future research.

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Genetic evidence of a redox-dependent systemic wound response via Hayan Protease-Phenoloxidase system inDrosophila

Cited by 106 publications

References 60 publications

Prophenoloxidase Activation Is Required for Survival to Microbial Infections in Drosophila

Prophenoloxidase Activation Is Required for Survival to Microbial Infections in Drosophila

The <b><i>Drosophila</i></b> Chitinase-Like Protein IDGF3 Is Involved in Protection against Nematodes and in Wound Healing

Methods to study Drosophila immunity

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