Isolation of a novel LPS-induced component of the ML superfamily in Ciona intestinalis

Vizzini, Aiti; Bonura, Angela; Longo, Valeria; Sanfratello, Maria Antonietta; Parrinello, Daniela; Cammarata, Matteo; Colombo, Piergiuseppe

doi:10.1016/j.dci.2015.06.018

Cited by 6 publications

(4 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The ascidian C. robusta possesses two Tlr-related genes presenting "hybrid" biological and immunological functionality of mammalian TLRs 19 , but LPS significantly activates only one of them, Tlr2; on the other hand, no MD-2 orthologue was found in the genome of C. robusta. The subtractive hybridization strategy allowed the identification of Npc2 mRNA in C. robusta, an homologue of ML superfamily components, preferentially expressed in haemocytes inside the vessel lumen of the pharynx 48 . This finding supports the hypothesis that the recognition of LPS by TLR4 through MD-2 binding may have been gained early during the evolution of vertebrates and that C. robusta may respond to LPS through a complex with alternative LPS sensors (i.e., the Ncp2 protein) developed before the differentiation of the MD-2 protein in vertebrates.…”

Section: Discussionmentioning

confidence: 99%

“…Relative expression was determined by dividing the normalised value of the target gene in each tissue by the normalised value obtained from the untreated tissue. To examine the time course of the response, LPS-treated ascidians (N = 4) were examined at incremental post-inoculation time points (1,2,4,8,12,24,48, and 72 h). Untreated ascidians (naïve) (N = 4) were used as controls.…”

Section: Gene Functional Enrichment Analysis and Immune Network Const...mentioning

confidence: 99%

See 1 more Smart Citation

Transcriptional and in silico analyses of MIF cytokine and TLR signalling interplay in the LPS inflammatory response of Ciona robusta

Arizza

Bonura

Paglia

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

The close phylogenetic relationship between Ciona robusta and vertebrates makes it a powerful model for studying innate immunity and the evolution of immune genes. To elucidate the nature and dynamics of the immune response, the molecular mechanisms by which bacterial infection is detected and translated into inflammation and how potential pattern recognition receptors (PRRs) are involved in pathogen recognition in tunicate C. robusta (formerly known as Ciona intestinalis), we applied an approach combining bacterial infections, next-generation sequencing, qRT-PCR, bioinformatics and in silico analyses (criteria of a p-value < 0.05 and FDR < 0.05). A STRING analysis indicated a functional link between components of the Tlr/MyD88-dependent signalling pathway (Tlr2, MyD88, and Irak4) and components of the Nf-κB signalling pathway (Nf-κB, IκBα, and Ikkα) (p-value < 0.05, FDR < 0.05). A qRT-PCR analysis of immune genes selected from transcriptome data revealed Mif as more frequently expressed in the inflammatory response than inflammation mediator or effector molecules (e.g., Il-17s, Tnf-α, Tgf-β, Mmp9, Tlrs, MyD88, Irak4, Nf-κB, and galectins), suggesting close interplay between Mif cytokines and Nf-κB signalling pathway components in the biphasic activation of the inflammatory response. An in silico analyses of the 3′-UTR of Tlr2, MyD88, IκBα, Ikk, and Nf-κB transcripts showed the presence of GAIT elements, which are known to play key roles in the regulation of immune gene-specific translation in humans. These findings provide a new level of understanding of the mechanisms involved in the regulation of the C. robusta inflammatory response induced by LPS and suggest that in C. robusta, as in humans, a complex transcriptional and post-transcriptional control mechanism is involved in the regulation of several inflammatory genes.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Gene Functional Enrichment Analysis and Immune Network Const...mentioning

confidence: 99%

Transcriptional and in silico analyses of MIF cytokine and TLR signalling interplay in the LPS inflammatory response of Ciona robusta

Arizza

Bonura

Paglia

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…In addition to this, ML family members are also known to play an important role by acting as agonists in dengue virus infection by modifying mosquito immunity (Jupatanakul et al, 2014). One of the invertebrate ML superfamily members identified in Ciona intestinalis homologous to the vertebrate npc2 gene and is induced after LPS inoculation (Vizzini et al, 2015). In the case of plant fungal pathogens, very few have been explored to characterize the functions of this superfamily.…”

Section: Introductionmentioning

confidence: 99%

Horizontal transfer of a conservednpc-2like effector gene in rust fungi that suppresses cell death in plants

Jaswal

Dubey

Kiran

et al. 2021

Preprint

View full text Add to dashboard Cite

ML/MD-2 is a conserved lipid/sterol-binding protein family having a role in sterol transfer and innate immunity in lower and higher eukaryotes. Here we report a genome-wide survey of this family, identifying 84 genes in 25 fungal and five oomycetes plant pathogen, having a different nutrition mode. All the fungal species were found to have varied numbers of family members, a distinctively substantial expansion of the ML gene family was observed in Rhizophagus irregularis (RI) with 33 genes. Our analysis also showed that NPC2 like proteins, a subfamily of ML domain superfamily, were not only restricted to animals and insect species but also present in plant fungal pathogens, including members of Clavicipitaceae, Pucciniacease, and Tremellaceae family. The phylogenetic analysis showed that these NPC2 like fungal proteins are more closely related to animals/insects than other fungal species. The molecular docking studies of these proteins with cholesterol and other derivatives indicate lipid-binding functional conservation across the animal and fungi kingdom. Further, the full length CDS of one of the npc2 like genes from Puccinia triticina (Pt5643) was PCR amplified and further characterized using various studies such as qRT-PCR, expression in onion epidermal cells, Nicotiana benthamiana for subcellular localization studies, yeast functional complementation, and expression studies. The mRNA abundance of Pt5643 was observed to be increased along with the infection progression and exhibits the highest expression at 5thday post-infection (dpi), suggesting its important role in the P. triticina infection cycle in wheat. The fluorescent confocal microscopy of transiently expressed YFP tagged Pt5643 in onion epidermal cells and N. benthamiana shows its location in cytoplasm and nucleus, indicating its involvement in the manipulation of host genes. The functional complementation of Pt5643 in npc2 mutant yeast showed its functional similarity to the eukaryotic npc2 gene. Further, the overexpression of Pt5643 also suppressed the BAX and H2O2 induced program cell death in N. benthamiana and yeast, respectively thus proving to be a novel horizontally transferred effector in rust fungal pathogens. Altogether the present study reports the novel function of fungal NPC2 like proteins playing a crucial role in host defense manipulation possibly through lipid binding/transport similar to animals.

show abstract

“…These responses can induce several immunological phenomena, including the expression of characteristic innate immune genes and a repertoire of innate effectors (e.g., TLRs, TNF, IL17s, TGF beta, complement components, and galectin-like) (Parrinello et al, 2008;Vizzini et al, 2016). Although a variety of different MAMPs induce Ciona TLRs, LPS does not activate TLR expression directly, as with TLR4 in vertebrates, suggesting the presence of alternative LPS sensors (Vizzini et al, 2015). In vertebrates, the development of the adaptive immune system is linked to the acquisition of the enzyme machinery encoded by Recombination Activating Genes (RAG) that provide the rearrangement of immunoglobulin (Ig) and TCR genes.…”

Section: Introductionmentioning

confidence: 99%

Ci8 short, a novel LPS-induced peptide from the ascidian Ciona intestinalis, modulates responses of the human immune system

et al. 2018

Self Cite

View full text Add to dashboard Cite

Isolation of a novel LPS-induced component of the ML superfamily in Ciona intestinalis

Cited by 6 publications

References 37 publications

Transcriptional and in silico analyses of MIF cytokine and TLR signalling interplay in the LPS inflammatory response of Ciona robusta

Transcriptional and in silico analyses of MIF cytokine and TLR signalling interplay in the LPS inflammatory response of Ciona robusta

Horizontal transfer of a conservednpc-2like effector gene in rust fungi that suppresses cell death in plants

Ci8 short, a novel LPS-induced peptide from the ascidian Ciona intestinalis, modulates responses of the human immune system

Contact Info

Product

Resources

About