Cloning and localization of MCdef, a defensin from Manila clams (Ruditapes philippinarum)

Adhya, Mausumi; Jeung, Hee-Do; Kang, Hyun-Sil; Choi, Kwang-Sik; Lee, Dong Sun; Cho, Moonjae

doi:10.1016/j.cbpb.2011.09.003

Cited by 21 publications

(14 citation statements)

References 26 publications

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“…Hence, understanding the host defense mechanisms of clams has become a primary concern. Previously, a defensin (MCdef) has been identified from Manila clam with the highest microbicidal activity against Streptococcus iniae and Staphylococcus aureus (Adhya et al, 2012). In this work, we report the characterization of a novel defensin from V. philippinarum, designated as VpDef.…”

Section: Contents Lists Available At Sciencedirectmentioning

confidence: 98%

“…Developmental and Comparative Immunology j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / d c i (Adhya et al, 2012;Bulet and Stocklin, 2005;Gueguen et al, 2006;Yang et al, 2000).…”

Section: Contents Lists Available At Sciencedirectmentioning

confidence: 99%

“…In recent years, multiple defensins have been identified from marine mollusks, such as Mytilus galloprovincialis (Hubert et al, 1996), Mytilus edulis (Charlet et al, 1996), Crassostrea virginica (Seo et al, 2005), Crassostrea gigas (Gonzalez et al, 2007;Gueguen et al, 2006), Haliotis discus discus (De Zoysa et al, 2010) and Venerupis philippinarum (Adhya et al, 2012).…”

Section: Contents Lists Available At Sciencedirectmentioning

confidence: 99%

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A defensin from clam Venerupis philippinarum: Molecular characterization, localization, antibacterial activity, and mechanism of action

Zhang

Yang

Wang

et al. 2015

Developmental & Comparative Immunology

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A B S T R A C TAntimicrobial peptides (AMPs) are important mediators of the primary host defense system against microbial invasion. In the present study, we cloned and characterized a member of the invertebrate defensin from the clam Venerupis philippinarum, designated VpDef. Amino acid sequence analysis showed that VpDef was similar to defensins from marine mollusks and ticks. In non-stimulated clams, RT-PCR and immunohistochemical analysis revealed that both VpDef mRNA and the encoding peptide were constitutively expressed in hemocytes and mantles, as well as in other major tissues. VpDef transcripts were significantly induced in hemocytes at different time intervals post Vibrio anguillarum infection. The recombinant VpDef (rVpDef) showed the highest activity against Gram-positive bacteria Micrococcus luteus and less effective to Gram-negative bacteria. In addition, incubation of rVpDef with M. luteus at 1 × and 3 × MIC could induce an obvious decrease of the membrane potential and notable changes of membrane permeability in a dose-dependent manner. Membrane integrity and bacterial viability analysis also revealed that rVpDef increased the membrane permeability of M. luteus and then resulted in cell death at 2 × and 10 × MIC. Overall, these results suggest that VpDef has an important function in host defense against invasive pathogens, probably killing microbes by inducing membrane lesions.

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Section: Contents Lists Available At Sciencedirectmentioning

confidence: 98%

Section: Contents Lists Available At Sciencedirectmentioning

confidence: 99%

Section: Contents Lists Available At Sciencedirectmentioning

confidence: 99%

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A defensin from clam Venerupis philippinarum: Molecular characterization, localization, antibacterial activity, and mechanism of action

Zhang

Yang

Wang

et al. 2015

Developmental & Comparative Immunology

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“…These types of tissueedistribution profiles suggested these defensins could respond promptly to bacterial challenge. However, it was reported that MCdef transcript was expressed at the highest level in adductor muscle of the Manila clam [32]; and the abalone defensin transcript was highly expressed in the mantle and hepatopancreas [19]. Thus, these results demonstrated that these defensins may play different immune roles in different tissues of these bivalves.…”

Section: Discussionmentioning

confidence: 73%

“…The expression of all Rpdefs was up-regulated remarkably after 12 h bacterial challenge compared to that of the control, indicating that the transcripts of Rpdefs were induced to fight against the bacteria. The up-regulation of defensin transcripts was also observed in disk abalone [19], Manila clam (MCdef) [32] and freshwater pearl mussel Hyriopsis schlegelii [44]. Then the expression levels of all Rpdefs except Rpdef4 returned to original level or were inhibited at 24 h and 48 h post challenge.…”

Section: Discussionmentioning

confidence: 85%

Molecular diversity and evolution of defensins in the manila clam Ruditapes philippinarum

Wang

Zhang

Yang

et al. 2015

Fish & Shellfish Immunology

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a b s t r a c tFour types of defensins were identified in Manila clam and designated as Rpdef1, Rpdef2, Rpdef3 and Rpdef4, which encoded a polypeptide of 49, 46, 45 and 42 amino acids, respectively. Sequence alignments indicated that Rpdef1 shared 46.9% identity with Rpdef2, 40.8% with Rpdef3, and 34.7% with Rpdef4. Analysis of transcript polymorphism showed that Rpdef3 accounted for about 60% frequency of Rpdefs occurrence in clams from three geographic origins (Dalian, Qingdao and Hangzhou). By quantitative real-time RT-PCR (qRT-PCR) analysis, the transcripts of Rpdefs were mainly detected in hemocytes and they responded sensitively to bacterial challenge in hemocytes. Evolutionary analysis indicated that all Rpdefs were under positive selection with positively selected basic amino acid residues detected in the C-terminal regions, which perhaps have a functional relevance by modifying the charge distribution of Rpdefs. The results also showed some lineages with dN/dS > 1, suggesting positive selection pressures existed in some lineages of phylogeny tree constructed by mollusk defensins. Overall, our results suggest that Rpdefs perhaps played important roles in host defense and positive selection is the major driving force in generating high diversity of defensins in the Manila clam.

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Antimicrobial Peptides in Marine Mollusks and their Potential Applications

Zoysa

2013

Marine Proteins and Peptides

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Cloning and localization of MCdef, a defensin from Manila clams (Ruditapes philippinarum)

Cited by 21 publications

References 26 publications

A defensin from clam Venerupis philippinarum: Molecular characterization, localization, antibacterial activity, and mechanism of action

A defensin from clam Venerupis philippinarum: Molecular characterization, localization, antibacterial activity, and mechanism of action

Molecular diversity and evolution of defensins in the manila clam Ruditapes philippinarum

Antimicrobial Peptides in Marine Mollusks and their Potential Applications

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