Antibacterial activity and immune responses of a molluscan macrophage expressed gene-1 from disk abalone, Haliotis discus discus.
The membrane-attack complex/perforin (MACPF) domain-containing proteins play an important role in the innate immune response against invading microbial pathogens. In the current study, a member of the MACPF domain-containing proteins, macrophage expressed gene-1 (MPEG1) encoding 730 amino acids with the theoretical molecular mass of 79.6 kDa and an isoelectric point (pI) of 6.49 was characterized from disk abalone Haliotis discus discus (AbMPEG1). We found that the characteristic MACPF domain (Val(131)-Tyr(348)) and transmembrane segment (Ala(669)-Ile(691)) of AbMPEG1 are located in the N- and C-terminal ends of the protein, respectively. Ortholog comparison revealed that AbMPEG1 has the highest sequence identity with its pink abalone counterpart, while sequences identities of greater than 90% were observed with MPEG1 members from other abalone species. Likewise, the furin cleavage site KRRRK was highly conserved in all abalone species, but not in other species investigated. We identified an intron-less genomic sequence within disk abalone AbMPEG1, which was similar to other mammalian, avian, and reptilian counterparts. Transcription factor binding sites, which are important for immune responses, were identified in the 5'-flanking region of AbMPEG1. qPCR revealed AbMPEG1 transcripts are present in every tissues examined, with the highest expression level occurring in mantle tissue. Significant up-regulation of AbMPEG1 transcript levels was observed in hemocytes and gill tissues following challenges with pathogens (Vibrio parahemolyticus, Listeria monocytogenes and viral hemorrhagic septicemia virus) as well as pathogen-associated molecular patterns (PAMPs: lipopolysaccharides and poly I:C immunostimulant). Finally, the antibacterial activity of the MACPF domain was characterized against Gram-negative and -positive bacteria using a recombinant peptide. Taken together, these results indicate that the biological significance of the AbMPEG1 gene includes a role in protecting disk abalone through the ability of AbMPEG1 to initiate an innate immune response upon pathogen invasion.
Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Research Center, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea.
S D N K
SD
Bathige
Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Research Center, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea.
Navaneethaiyer
N
Umasuthan
Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Research Center, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea.
Ilson
I
Whang
Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Research Center, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea.
Bong-Soo
BS
Lim
Ocean and Fisheries Research Institute, Jeju Special Self-Governing Province 699-915, Republic of Korea.
Seung Hwan
SH
Won
Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Self-Governing Province 690-756, Republic of Korea; Fish Vaccine Research Center, Jeju National University, Jeju Special Self-Governing Province 690-756, Republic of Korea. Electronic address: jehee@jejunu.ac.kr.
Jehee
J
Lee
D000704:Analysis of Variance; D000818:Animals; D001419:Bacteria / Q000276:immunology*; D001483:Base Sequence; D049851:Gastropoda / Q000276:immunology* / Q000382:microbiology*; D020869:Gene Expression Profiling; D007113:Immunity, Innate / Q000276:immunology*; D008264:Macrophages / Q000378:metabolism*; D008969:Molecular Sequence Data; D054353:Perforin / Q000378:metabolism*; D011487:Protein Conformation; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D054353:Perforin
Antibacterial activity; Genomic structure; Haliotis discus discus; Immune response; Macrophage expressed gene