title: Immune response of apoptosis-related cysteine peptidases from the red abalone Haliotis rufescens (HrCas8 and HrCas3): molecular characterization and transcription expression.
abstract: Caspases play an important role in the different stages of programmed cell death, or apoptosis, which has been related to the immune response in multicellular organisms. The present study characterized an initiator caspase (HrCas8) and an effector caspase (HrCas3) from the red abalone Haliotis rufescens using the RACE method and qPCR analysis. HrCas8 showed a complete sequence of 2529 base pairs (bp) with an open-reading frame (ORF) of 1911 bp, a 5'UTR of 201 bp, and a 3'UTR of 417 bp. The estimated molecular mass for the 636 amino acids from HrCas8 was 71.5 kDa with an isoelectric point of 6.2. The HrCas8 sequence had two death-effector domains (DEDs) and the subunits p20 and p10, in addition to an active site characteristic of cysteine proteins. Meanwhile, the effector caspase HrCas3 showed a complete sequence of 1404 bp, a 5'UTR of 82 bp, and a 3'UTR of 574 bp. The ORF of this caspase had 747 bp that coded for 248 residues. Moreover, the predicted molecular mass of HrCas3 was 29.4 kDa; the theoretical isoelectric point was 5.70, and the sequence evidenced a conserved caspase recruitment domain (CARD). The distribution of the caspases in distinct tissues revealed that HrCas8 was principally expressed in the hemolymph, while HrCas3 had a higher expression in the gills. A basal level of expression was found for both caspases in muscle tissue. The immune response of caspases in H. rufescens was evaluated through an injection of Vibrio anguillarum. The results showed an increase in the transcription of HrCas8 post-challenge, as well as an activation of HrCas3, which together suggest the initiation of apoptosis as a response to bacterial infection in H. rufescens.
authors.affiliation: Laboratory of Biotechnology and Aquatic Genomics, Interdisciplinary Center for Aquaculture Research (INCAR), University of Concepción, P.O. Box 160-C, Concepción, Chile.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Chávez-Mardones
authors.affiliation: Laboratory of Biotechnology and Aquatic Genomics, Interdisciplinary Center for Aquaculture Research (INCAR), University of Concepción, P.O. Box 160-C, Concepción, Chile. Electronic address: crisgallardo@udec.cl.
authors.forename: Cristian
authors.initials: C
authors.lastname: Gallardo-Escárate
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D053148:Caspase 3 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D053181:Caspase 8 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D020224:Expressed Sequence Tags; D015971:Gene Expression Regulation, Enzymologic; D005880:Gills / Q000378:metabolism; D006458:Hemolymph / Q000378:metabolism; D007113:Immunity, Innate; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D012908:Snails / Q000201:enzymology / Q000235:genetics / Q000276:immunology; D059467:Transcriptome; D014733:Vibrio / Q000502:physiology
chemical_list: D053148:Caspase 3; D053181:Caspase 8
keywords: Apoptosis; Gene expression; Haliotis rufescens; HrCas3; HrCas8