Abstract
PGRP-LC is a significant pattern recognition receptor of the insect innate immune system that can recognize peptidoglycans and activate immune signaling pathways regulating the expression and release of antimicrobial peptides against infection. We for the first time analyzed the phylogenetic tree, purification and structure of bumblebee PGRP-LC. The results showed high conservation of bumblebee PGRP-LC among the 16 bumblebee species, and further phylogenetic analysis showed that the PGRP-LC phylogeny of different subgenera (Subterraneobombus, Megabombus, Melanobombus, Bombus) is consistent with that of the COI gene. Additionally, the phylogeny of PGRP-LCs among Bombus, Apis and the solitary bee Megachile rotundata coincides with the sociality evolution of bees. Moreover, bumblebee PGRP-LC (Bl-PGRP-LC) shares the Drosophila PGRP-LCx and PGRP-LCa topology, retaining conserved disulfide bonds and 80% binding residues involved in the interaction between TCT and PGRP-LCx. Therefore, Bl-PGRP-LC might share some similar binding characteristics with Drosophila PGRP-LCx. In addition, Bl-PGRP-LC has shorter β5 and β1 sheets, longer β2, β3, and β4 sheets and a shallow binding groove. To determine the characteristics of Bl-PGRP-LC, high-purity PGRP-LC inclusion bodies, soluble GST-tag Bl-PGRP-LC fusion protein and soluble pure Bl-PGRP-LC were obtained in vitro. The results will be helpful for further study of the function and structure of Bl-PGRP-LC.
Highlights
Bumblebees are important pollinators and play a key role in agricultural and natural ecosystems (An et al, 2014; P Williams, Colla, & Xie, 2009)
Based on alignment to the B. terrestris and B. impatients genomes, Peptidoglycan recognize protein LC (PGRP-LC) consists of four exons (Exon 1: 6645776 nt-6646784 nt, Exon 2: 6647933 nt-6648518 nt, Exon 3: 6649718 nt-6649876 nt, Exon 4: 6650035 nt-6650667 nt), with a length of 2742 bp (Fig 1B)
The GST-tag was successfully cleaved by the PSP enzyme, and pure GST-tag and BlPGRP-LC bands were visualized by SDS-PAGE (Fig 5C)
Summary
Bumblebees are important pollinators and play a key role in agricultural and natural ecosystems (An et al, 2014; P Williams, Colla, & Xie, 2009). Resistance to pathogens occurs through innate immunity, and responses to pathogens are mainly triggered by insect pattern recognition receptors (PRRs) recognition of pathogen-associated molecular pattern (PAMPs), such as lipopolysaccharide, lipteichoic acid, peptidoglycans (PGNs), and β-1,3- dextran, which are produced by most pathogenic microorganisms (Yu, Zhu, Ma, Fabrick, & Kanost, 2002). PGNs are among the most important constituents of gram-negative and -positive bacterial cell walls and can be divided into Dap-type PGN and Lys-type PGN according to differences in the third amino acid of the short peptide (Schleifer & Kandler, 1972). Dap-type PGN is largely found in all gram-negative bacteria and some grampositive bacteria, e.g., Bacillus and Clostridium spp., whereas Lys-type PGN is mainly found in gram-positive bacteria and is absent in gram-negative bacteria (Kurata, 2014)
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