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Title: Identification of SHIP-1 and SHIP-2 homologs in channel catfish, Ictalurus punctatus

Author
item TAYLOR, ERIN - University Of Mississippi Medical Center
item NAYAK, DEEPAK - Washington University
item Quiniou, Sylvie
item BENGTEN, EVA - University Of Mississippi Medical Center
item WILSON, MELANIE - University Of Mississippi Medical Center

Submitted to: Developmental and Comparative Immunology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/24/2015
Publication Date: 7/1/2015
Citation: Taylor, E.B., Nayak, D.K., Quiniou, S., Bengten, E., Wilson, M. 2015. Identification of SHIP-1 and SHIP-2 homologs in channel catfish, Ictalurus punctatus. Developmental and Comparative Immunology. 51(1):79-87.

Interpretive Summary: Src homology domain 2 (SH2) domain-containing inositol 5’-phosphatases (SHIP) proteins are important enzymes that modulate immune cell signaling pathways. In this study, catfish SHIP-1 and SHIP-2 genes were identified. Structurally these genes contain all the features and conserved motifs that mediate SHIP-1 and SHIP-2 function in mammals. Also their gene expression pattern is similar to the pattern observed with mammalian SHIP genes. Importantly, commercially available anti-SHIP antibodies were shown to recognize catfish SHIP proteins. Since little is known about SHIP protein function in fish, these antibodies can be now be used to study immune protein signaling in catfish and likely as well in other fish.

Technical Abstract: Src homology domain 2 (SH2) domain-containing inositol 5’-phosphatases (SHIP) proteins have diverse roles in signal transduction. SHIP-1 and SHIP-2 homologs were identified in channel catfish, Ictalurus punctatus, based on sequence homology to murine and human SHIP sequences. Full-length cDNAs for catfish SHIP-1 and SHIP-2 (IpSHIP-1 and IpSHIP-2) were obtained using 5' and 3' RACE protocols. Catfish SHIP molecules share a high degree of sequence identity to their respective SHIP sequences from diverse taxa and both are encoded by single copy genes. IpSHIP-1 and IpSHIP-2 transcripts were expressed in all catfish tissues analyzed except for skin, and IpSHIP-1 message was more abundant than IpSHIP-2 message in lymphoid tissues. Catfish clonal B, cytotoxic T, and macrophage cell lines also expressed message for both molecules. IpSHIP-1 and IpSHIP-2 SH2 domains were expressed as recombinant proteins and were both found to be bound by cross-reacting rabbit anti-mouse SHIP-1 pAb. The anti-mouse SHIP-1 pAb also reacted with cell lysates from the cytotoxic T cell lines, TS32.15 and TS32.17 as well as 42TA macrophages.