Dataset Information

CDNA and Gene Structure of MytiLec-1, A Bacteriostatic R-Type Lectin from the Mediterranean Mussel (Mytilus galloprovincialis).

ABSTRACT: MytiLec is an ?-d-galactose-binding lectin with a unique primary structure isolated from the Mediterranean mussel (Mytilus galloprovincialis). The lectin adopts a ?-trefoil fold that is also found in the B-sub-unit of ricin and other ricin-type (R-type) lectins. We are introducing MytiLec(-1) and its two variants (MytiLec-2 and -3), which both possess an additional pore-forming aerolysin-like domain, as members of a novel multi-genic "mytilectin family" in bivalve mollusks. Based on the full length mRNA sequence (911 bps), it was possible to elucidate the coding sequence of MytiLec-1, which displays an extended open reading frame (ORF) at the 5' end of the sequence, confirmed both at the mRNA and at the genomic DNA sequence level. While this extension could potentially produce a polypeptide significantly longer than previously reported, this has not been confirmed yet at the protein level. MytiLec-1 was revealed to be encoded by a gene consisting of two exons and a single intron. The first exon comprised the 5'UTR and the initial ATG codon and it was possible to detect a putative promoter region immediately ahead of the transcription start site in the MytiLec-1 genomic locus. The remaining part of the MytiLec-1 coding sequence (including the three sub-domains, the 3'UTR and the poly-A signal) was included in the second exon. The bacteriostatic activity of MytiLec-1 was determined by the agglutination of both Gram-positive and Gram-negative bacteria, which was reversed by the co-presence of ?-galactoside. Altogether, these data support the classification of MytiLec-1 as a member of the novel mytilectin family and suggest that this lectin may play an important role as a pattern recognition receptor in the innate immunity of mussels.

SUBMITTER: Hasan I

PROVIDER: S-EPMC4882566 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature

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Publications

cDNA and Gene Structure of MytiLec-1, A Bacteriostatic R-Type Lectin from the Mediterranean Mussel (Mytilus galloprovincialis).

Hasan Imtiaj I Gerdol Marco M Fujii Yuki Y Rajia Sultana S Koide Yasuhiro Y Yamamoto Daiki D Kawsar Sarkar M A SM Ozeki Yasuhiro Y

Marine drugs 20160511 5

MytiLec is an α-d-galactose-binding lectin with a unique primary structure isolated from the Mediterranean mussel (Mytilus galloprovincialis). The lectin adopts a β-trefoil fold that is also found in the B-sub-unit of ricin and other ricin-type (R-type) lectins. We are introducing MytiLec(-1) and its two variants (MytiLec-2 and -3), which both possess an additional pore-forming aerolysin-like domain, as members of a novel multi-genic "mytilectin family" in bivalve mollusks. Based on the full len ...[more]

PMID: 27187419

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Project description:BACKGROUND: Sessile bivalves of the genus Mytilus are suspension feeders relatively tolerant to a wide range of environmental changes, used as sentinels in ecotoxicological investigations and marketed worldwide as seafood. Mortality events caused by infective agents and parasites apparently occur less in mussels than in other bivalves but the molecular basis of such evidence is unknown. The arrangement of Mytibase, interactive catalogue of 7,112 transcripts of M. galloprovincialis, offered us the opportunity to look for gene sequences relevant to the host defences, in particular the innate immunity related genes. RESULTS: We have explored and described the Mytibase sequence clusters and singletons having a putative role in recognition, intracellular signalling, and neutralization of potential pathogens in M. galloprovincialis. Automatically assisted searches of protein signatures and manually cured sequence analysis confirmed the molecular diversity of recognition/effector molecules such as the antimicrobial peptides and many carbohydrate binding proteins. Molecular motifs identifying complement C1q, C-type lectins and fibrinogen-like transcripts emerged as the most abundant in the Mytibase collection whereas, conversely, sequence motifs denoting the regulatory cytokine MIF and cytokine-related transcripts represent singular and unexpected findings. Using a cross-search strategy, 1,820 putatively immune-related sequences were selected to design oligonucleotide probes and define a species-specific Immunochip (DNA microarray). The Immunochip performance was tested with hemolymph RNAs from mussels injected with Vibrio splendidus at 3 and 48 hours post-treatment. A total of 143 and 262 differentially expressed genes exemplify the early and late hemocyte response of the Vibrio-challenged mussels, respectively, with AMP trends confirmed by qPCR and clear modulation of interrelated signalling pathways. CONCLUSIONS: The Mytibase collection is rich in gene transcripts modulated in response to antigenic stimuli and represents an interesting window for looking at the mussel immunome (transcriptomes mediating the mussel response to non-self or abnormal antigens). On this basis, we have defined a new microarray platform, a mussel Immunochip, as a flexible tool for the experimental validation of immune-candidate sequences, and tested its performance on Vibrio-activated mussel hemocytes. The microarray platform and related expression data can be regarded as a step forward in the study of the adaptive response of the Mytilus species to an evolving microbial world.

Project description:BackgroundMussels provide many ecosystem services as habitat, food, water filtration and recreational fishing. However, mussels are vulnerable to anthropogenic pressures such as harvesting or trampling, among others. In this frame, it would be paramount to engage society in marine conservation and improving its awareness about environmental policies. The first step lies in properly assessing what is the perception and concerns of society about marine ecosystems. Our study aims to fill this gap by examining public perception of services provided by Mytilus galloprovincialis, its state of conservation and the factors (including anthropogenic activities) shaping mussel beds.MethodsThis study is based on a face-to-face survey consisting of seven open-ended and seven multiple-choice questions of 404 people conducted in 2019 at different shores in the North Portuguese coast. The influence of respondent profile in terms of age, education, gender and place of residence was also assessed.ResultsMost of the participants in our survey (74%) considered that mussels contributed to human well-being and life quality; however, only 31% considered that mussels provide us with many benefits. Regarding the perceived state of mussel services, most of the respondents asserted that mussel services (purification of seawater, habitat, food for other species) worsened in the last 10 years. In contrast, the service as human food was perceived as in an identical state and scientific and traditional knowledge was the only service perceived in a better state. Concerning the state of mussel beds, most of the participants perceived it as good (45%) but a similar percentage (41%) asserted ignoring it. When considering the influence of different factors on mussel beds, only environmental management was considered as having a positive impact by a higher percentage of respondents. The majority of the participants considered that factors included in the questionnaire contributed to worsen mussel beds, ranging between 51% for coastal erosion and 90% for pollution. Education level and age were the main socio-economic factors driving public awareness about the importance of mussel services, its state of conservation and the factors shaping mussel beds.DiscussionResults showed that perception about the importance of mussels for human well-being and the quantity of delivered benefits increased with the education level. Moreover, older people perceived human food as the most important service offered by mussels. Therefore, our results suggest that mussels are mainly known as food resource; however, most of the people ignore their relevant ecological role and the many other benefits that mussels provide. Thus, it is necessary to actively engage society about importance of mussel beds. As M. galloprovincialis is a relevant economic resource, our data could improve the diffusion of knowledge among citizens, stakeholders and scientists, contributing to its sustainability.

Dataset Information

CDNA and Gene Structure of MytiLec-1, A Bacteriostatic R-Type Lectin from the Mediterranean Mussel (Mytilus galloprovincialis).

Publications

cDNA and Gene Structure of MytiLec-1, A Bacteriostatic R-Type Lectin from the Mediterranean Mussel (Mytilus galloprovincialis).

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