Molecular and functional characterization of gilthead seabream Sparus aurata caspase-1: the first identification of an inflammatory caspase in fish.
Caspases are a family of cysteine proteases that fulfil critical roles in mammalian apoptosis and in the proteolytic activation of cytokines. In humans, the caspase family includes 13 members whose functions seem to correlate with their phylogenetic relationship. They are classified into two main groups, the cell death (apoptotic) and the inflammatory caspases. Caspase-1 is the best characterized inflammatory caspase and is responsible for the processing of interleukin-1beta (IL-1beta), IL-18 and IL-33. Despite the importance of caspase-1 in inflammation, no information is available on the presence and activity of this enzyme in fish. In this study, we cloned a caspase-1-like gene from the bony fish gilthead seabream (Sparus aurata L.) which shows a conserved N-terminal caspase-recruitment domain (CARD) and a C-terminal caspase catalytic domain. The seabream caspase-1 gene was expressed in 1 day post-hatching larvae and its mRNA levels increased throughout development. In adult fish, caspase-1 was found to be constitutively expressed in all immune tissues analyzed and, unexpectedly, infection of fish and stimulation of professional phagocytes in vitro decreased its mRNA levels. It was also demonstrated that the recombinant seabream caspase-1 ectopically expressed in HEK293 cells was able to cleave a caspase-1 specific substrate, this activity being enhanced upon activation of the rat P2X7 receptor with BzATP. Finally, seabream fibroblast cell line SAF-1 and primary leukocytes showed endogenous caspase-1 activity, which was almost completely inhibited by a caspase-1 specific inhibitor.
López-Castejón
Gloria
G
Department of Cell Biology, Faculty of Biology, University of Murcia, 30100 Murcia, Spain.
Sepulcre
M Pilar
MP
Mulero
Iván
I
Pelegrín
Pablo
P
Meseguer
José
J
Mulero
Victoriano
V
D000595:Amino Acid Sequence; D000818:Animals; D001424:Bacterial Infections / Q000201:enzymology; D020170:Caspase 1 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D015536:Down-Regulation / Q000235:genetics; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007249:Inflammation; D007962:Leukocytes / Q000201:enzymology; D008969:Molecular Sequence Data; D010586:Phagocytes / Q000201:enzymology; D010802:Phylogeny; D011994:Recombinant Proteins / Q000378:metabolism; D021541:Sea Bream / Q000235:genetics* / Q000382:microbiology; D017386:Sequence Homology, Amino Acid
D011994:Recombinant Proteins; D020170:Caspase 1