Cloning and characterization of a novel activating Ly49 closely related to Ly49A

• Published in: The Journal of immunology (1950) 1999-11, Vol.163 (9), p.4931-4938
• Main Authors: Makrigiannis, A P, Gosselin, P, Mason, L H, Taylor, L S, McVicar, D W, Ortaldo, J R, Anderson, S K
• Format: Article
• Language: English
• Subjects: Adaptor Proteins, Signal Transducing; Amino Acid Sequence; Animals; Antibodies, Monoclonal - metabolism; Antigen-Antibody Reactions; Antigens, Ly - chemistry; Antigens, Ly - genetics; Antigens, Ly - immunology; Antigens, Ly - metabolism; Base Sequence; Calcium Signaling - immunology; Carrier Proteins - genetics; Cloning, Molecular; DNA, Complementary - isolation & purification; Humans; Killer Cells, Natural - immunology; Killer Cells, Natural - metabolism; Lectins, C-Type; Lymphocyte Activation - immunology; Membrane Proteins - genetics; Mice; Mice, Inbred Strains; Molecular Sequence Data; NK Cell Lectin-Like Receptor Subfamily A; Phosphorylation; Polymerase Chain Reaction; Receptors, Immunologic - chemistry; Receptors, Immunologic - genetics; Receptors, Immunologic - immunology; Receptors, Immunologic - metabolism; Receptors, Immunologic - physiology; Receptors, NK Cell Lectin-Like
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.163.9.4931

The majority of the known Ly49 family members have been isolated from either C57BL/6 (B6) or BALB/c mice. Interestingly, the anti-Ly49 Ab reactivities observed in 129/J mice are different from those of B6 mice. Furthermore, immunoprecipitation of 129/J NK cell lysates with YE1/32 and YE1/48, Abs specific for the inhibitory Ly49A in B6, resulted in detection of the activation-associated DAP12 molecule. These results indicated a need for a more detailed study of this strain. Therefore, a cloning strategy was devised to isolate Ly49 cDNAs from 129/J mice. An immunoreceptor tyrosine-based inhibitory motif-containing, Ly49D-related clone was discovered that we have named Ly49O, and one immunoreceptor tyrosine-based inhibitory motif-lacking, Ly49A-related clone was discovered that we have named Ly49P. No anti-Ly49 mAb reacted with Ly49O, whereas the molecule encoded by the Ly49P cDNA was found to react with YE1/32 and YE1/48. Ly49P was found to associate with mouse DAP12, and Ab-mediated cross-linking of Ly49P resulted in mouse DAP12 phosphorylation and Ca2+ mobilization, indicating that Ly49P is a competent activation receptor. Ly49P, therefore, represents a novel member of the Ly49 activating receptor subfamily.