Abstract
Endogenous antimicrobial peptides (AMPs) are evolutionarily ancient factors of innate immunity, which are produced by all multicellular organisms and play a key role in their protection against infection. Red king crab (Paralithodes camtschaticus), also called Kamchatka crab, is widely distributed and the best known species of all king crabs belonging to the family Lithodidae. Despite their economic importance, the genetic resources of king crabs are scarcely known and no full-genome sequences are available to date. Therefore, analysis of the red king crab transcriptome and identification and characterization of its AMPs could potentially contribute to the development of novel antimicrobial drug candidates when antibiotic resistance has become a global health threat. In this study, we sequenced the P. camtschaticus transcriptomes from carapace, tail flap and leg tissues using an Illumina NGS platform. Libraries were systematically analyzed for gene expression profiles along with AMP prediction. By an in silico approach using public databases we defined 49 cDNAs encoding for AMP candidates belonging to diverse families and functional classes, including buforins, crustins, paralithocins, and ALFs (anti-lipopolysaccharide factors). We analyzed expression patterns of 27 AMP genes. The highest expression was found for Paralithocin 1 and Crustin 3, with more than 8,000 reads. Other paralithocins, ALFs, crustins and ubiquicidins were among medium expressed genes. This transcriptome data set and AMPs provide a solid baseline for further functional analysis in P. camtschaticus. Results from the current study contribute also to the future application of red king crab as a bio-resource in addition to its being a known seafood delicacy.
Highlights
Endogenous antimicrobial peptides (AMPs) are evolutionarily ancient factors of innate immunity of multicellular organisms, and play a key role in their protection against infection[1,2]
Due to the lack of adaptive immunity which is present in vertebrates, invertebrates use relatively simple defense strategies that mainly rely on AMPs and innate immunity mechanisms, such as protective barriers, toxic molecules, and phagocytic cells that ingest and destroy invading microorganisms and larger parasites[10]
The de novo transcriptomes of four types of tissues of two individuals of the red king crab P. camtschaticus were sequenced on Illumina HiSeq 3/4,000 using two lanes and 2 × 150 bp. This resulted in a draft gene set of 384,854 transcripts and 274,917 uni-genes of red king crab
Summary
Endogenous antimicrobial peptides (AMPs) are evolutionarily ancient factors of innate immunity of multicellular organisms, and play a key role in their protection against infection[1,2]. AMPs were commonly considered to be a primitive mechanism of immunity and have been extensively studied in insects and other non-vertebrate organisms. AMP-encoding genes are either constitutively expressed or rapidly transcribed upon induction in eukaryotes by invading microbes and their products. These peptides are classified into families with distinct properties based on their amino acid sequences, number of cysteine residues, and spacing[9]. AMPs increase the permeability of the cell membrane, thereby enhancing the effect of traditionally used antibiotics and can be used in combination with t hem[11,13]
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