Short-Form Bomanins Mediate Humoral Immunity in &lt;b&gt;&lt;i&gt;Drosophila&lt;/i&gt;&lt;/b&gt;

Lindsay, Scott A.; Lin, Samuel; Wasserman, Steven A.

doi:10.1159/000489831

Cited by 57 publications

(99 citation statements)

References 21 publications

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“…Plasmids expressing dso1 or dso2 transcripts from the pBomS3 promoter were made using methods previously described (23). Briefly, the BomS3 gene promoter was placed 5 ′ to the ORF encoding either Dso1 or Dso2.…”

Section: Fly Husbandry and Strain Generationmentioning

confidence: 99%

“…For the induction of the Toll response, heat-killed Micrococcus luteus was prepared as previously described (23).…”

Section: Microbial Culturesmentioning

confidence: 99%

“…After Toll induction with heat-killed M. luteus, flies were incubated at 29 • C for 24 h, after which hemolymph was collected via capillary as previously described (23). Hemolymph in 0.1% trifluoroacetic acid/50% acetonitrile was mixed 1:1 with Universal Matrix (Sigma-Aldrich).…”

Section: Maldi-tofmentioning

confidence: 99%

“…Pioneering mass spectrometry experiments by Bulet et al identified two dozen peptide IMs (immune-induced molecules) that accumulate in Drosophila hemolymph upon induction of the innate immune response, principally the Toll pathway (13,16). Among these, the Bomanins have been found to play an essential role against a broad range of pathogens (21,23) while several, including the 15 aa long IM4 and 24 aa long IM14, have unknown functions. Based on our demonstration of defensive functions for these peptides, we have renamed them Daisho1 and Daisho2, for daisho, the Japanese term for a matched pair of samurai swords, one short and one long.…”

Section: Generation Of Flies Null For the Daisho Gene Pairmentioning

confidence: 99%

“…Alternatively, survival might depend only on total dosage for the two genes, with loss of either dropping expression below the threshold required. To distinguish between these models, we generated transgenes placing each ORF under control of pBomS3, shown previously to be strongly Toll-responsive promoter (23), and then assayed the transgenes for rescue of dso1 or dso2. As shown in Table 1, pBomS3-driven dso1 rescued dso1, improving the median survival from 46 to 93 h (p < 0.0001).…”

Section: Daisho1 and Daisho2 Are Each Required For Defensementioning

confidence: 99%

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The Daisho Peptides Mediate Drosophila Defense Against a Subset of Filamentous Fungi

Cohen

Lindsay

et al. 2020

Front. Immunol.

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Fungal infections, widespread throughout the world, affect a broad range of life forms, including agriculturally relevant plants, humans, and insects. In defending against fungal infections, the fruit fly Drosophila melanogaster employs the Toll pathway to induce a large number of immune peptides. Some have been investigated, such as the antimicrobial peptides (AMPs) and Bomanins (Boms); many, however, remain uncharacterized. Here, we examine the role in innate immunity of two related peptides, Daisho1 and Daisho2 (formerly IM4 and IM14, respectively), found in hemolymph following Toll pathway activation. By generating a CRISPR/Cas9 knockout of both genes, daisho, we find that the Daisho peptides are required for defense against a subset of filamentous fungi, including Fusarium oxysporum, but not other Toll-inducible pathogens, such as Enterococcus faecalis and Candida glabrata. Analysis of null alleles and transgenes revealed that the two daisho genes are each required for defense, although their functions partially overlap. Generating and assaying a genomic epitope-tagged Daisho2 construct, we detected interaction in vitro of Daisho2 peptide in hemolymph with the hyphae of F. oxysporum. Together, these results identify the Daisho peptides as a new class of innate immune effectors with humoral activity against a select set of filamentous fungi.

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Section: Fly Husbandry and Strain Generationmentioning

confidence: 99%

“…For the induction of the Toll response, heat-killed Micrococcus luteus was prepared as previously described (23).…”

Section: Microbial Culturesmentioning

confidence: 99%

Section: Maldi-tofmentioning

confidence: 99%

Section: Generation Of Flies Null For the Daisho Gene Pairmentioning

confidence: 99%

Section: Daisho1 and Daisho2 Are Each Required For Defensementioning

confidence: 99%

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The Daisho Peptides Mediate Drosophila Defense Against a Subset of Filamentous Fungi

Cohen

Lindsay

et al. 2020

Front. Immunol.

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The evolutionary novelty of insect defensins: from bacterial killing to toxin neutralization

Gao,

Zhu

2024

Cell. Mol. Life Sci.

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Insect host defense comprises two complementary dimensions, microbial killing-mediated resistance and microbial toxin neutralization-mediated resilience, both jointly providing protection against pathogen infections. Insect defensins are a class of effectors of innate immunity primarily responsible for resistance to Gram-positive bacteria. Here, we report a newly originated gene from an ancestral defensin via genetic deletion following gene duplication in Drosophila virilis, which confers an enhanced resilience to Gram-positive bacterial infection. This gene encodes an 18-mer arginine-rich peptide (termed DvirARP) with differences from its parent gene in its pattern of expression, structure and function. DvirARP specifically expresses in D. virilis female adults with a constitutive manner. It adopts a novel fold with a 310 helix and a two CXC motif-containing loop stabilized by two disulfide bridges. DvirARP exhibits no activity on the majority of microorganisms tested and only a weak activity against two Gram-positive bacteria. DvirARP knockout flies are viable and have no obvious defect in reproductivity but they are more susceptible to the DvirARP-resistant Staphylococcus aureus infection than the wild type files, which can be attributable to its ability in neutralization of the S. aureus secreted toxins. Phylogenetic distribution analysis reveals that DvirARP is restrictedly present in the Drosophila subgenus, but independent deletion variations also occur in defensins from the Sophophora subgenus, in support of the evolvability of this class of immune effectors. Our work illustrates for the first time how a duplicate resistance-mediated gene evolves an ability to increase the resilience of a subset of Drosophila species against bacterial infection.

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Genome-wide transcriptional effects of deletions of sulphur metabolism genes in Drosophila melanogaster

et al. 2020

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In recent years, the gasotransmitter hydrogen sulphide (H 2 S), produced by the transsulphuration pathway, has been recognized as a biological mediator playing an important role under normal conditions and in various pathologies in both eukaryotes and prokaryotes. The transsulphuration pathway (TSP) includes the conversion of homocysteine to cysteine following the breakdown of methionine. In Drosophila melanogaster and other eukaryotes, H 2 S is produced by cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE), and 3-mercaptopyruvate sulphurtransferase (MST). In the experiments performed in this study, we were able to explore the CRISPR/Cas9 technique to obtain single and double deletions in homozygotes of these three major genes responsible for H 2 S production in Drosophila melanogaster . In most cases, the deletion of one studied gene does not result in the compensatory induction of two other genes responsible for H 2 S production. Transcriptomic studies demonstrated that the deletions of the above CBS and CSE genes alter genome expression to different degrees, with a more pronounced effect being exerted by deletion of the CBS gene. Furthermore, the double deletion of both CBS and CSE resulted in a cumulative effect on transcription in the resulting strains. Overall, we found that the obtained deletions affect numerous genes involved in various biological pathways. Specifically, genes involved in the oxidative reduction process, stress-response genes, housekeeping genes, and genes participating in olfactory and reproduction are among the most strongly affected. Furthermore, characteristic differences in the response to the deletions of the studied genes are apparently organ-specific and have clear-cut sex-specific characteristics. Single and double deletions of the three genes responsible for the production of H 2 S helped to elucidate new aspects of the biological significance of this vital physiological mediator.

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Short-Form Bomanins Mediate Humoral Immunity in <b><i>Drosophila</i></b>

Cited by 57 publications

References 21 publications

The Daisho Peptides Mediate Drosophila Defense Against a Subset of Filamentous Fungi

The Daisho Peptides Mediate Drosophila Defense Against a Subset of Filamentous Fungi

The evolutionary novelty of insect defensins: from bacterial killing to toxin neutralization

Genome-wide transcriptional effects of deletions of sulphur metabolism genes in Drosophila melanogaster

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