RESUMEN
A cDNA clone for an IgG-binding Fc receptor, rtFc gamma R alpha, of the rat natural killer cell line CRNK-16 is characterized here. This clone encodes an Fc gamma receptor as shown by the ability of cDNA-transfected COS cells to rosette IgG-coated sheep erythrocytes. The rtFc gamma R alpha is exceptionally homologous to the mouse moFc gamma R alpha, with 77% protein sequence identity and 71% nucleic acid identity overall. The transmembrane region of the rtFc gamma R alpha contains the sequence Leu-Phe-Ala-Val-Asp-Thr-Gly-Leu, which is present in the membrane sequences of four other Fc receptors including mouse Fc gamma R alpha, human Fc gamma RIII-2, and the Fc epsilon R alpha subunits of the rat and human high-affinity IgE-binding receptors. Also, the rtFc gamma R alpha cytoplasmic domain exhibits specific homology to other receptors derived from natural killer cells, human Fc gamma RIII-2 and mouse Fc gamma R alpha. However, the rtFc gamma R alpha cDNA clone is complementary to at least two different-sized mRNAs expressed by CRNK-16 cells, contrasting the single Fc gamma R-related mRNA species expressed by human and mouse natural killer cells. These rat mRNAs are homologous to both the 5' and the 3' end of the cDNA clone, suggesting that they may be (i) splice variants of one transcript or (ii) products of different but highly related genes.
Asunto(s)
Antígenos de Diferenciación/genética , Células Asesinas Naturales/inmunología , Receptores Fc/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Northern Blotting , Línea Celular , Clonación Molecular , ADN/genética , Biblioteca de Genes , Humanos , Inmunoglobulina G/inmunología , Datos de Secuencia Molecular , Sondas de Oligonucleótidos , Poli A/genética , Poli A/aislamiento & purificación , ARN/genética , ARN/aislamiento & purificación , ARN Mensajero , Ratas , Receptores de IgG , Mapeo Restrictivo , Formación de Roseta , Homología de Secuencia de Ácido Nucleico , TransfecciónRESUMEN
The cDNA for the common beta Mac-1 subunit (CD11b) of the mouse LFA-1/Mac-1/p150,95 group of leukocyte cell adhesion receptors, formally designated integrin beta 2, has been cloned and sequenced. Clones were isolated from cDNA libraries made from J774 macrophage and WEHI-3B myelomonocytic tumor cells which express this subunit as a component of the macrophage activation antigen 1 (Mac-1), also known as complement receptor type 3 (CR3). This subunit is expressed as a single, abundant mRNA species approximately 2.7 kilobase (kb) in size. The 2422 base pair (bp) cDNA sequence obtained codes for a 771 amino acid protein organized with leader, extracellular, transmembrane, and cytoplamic domains of 23, 680, 23, and 46 amino acids, respectively, yielding an 82,700 mature protein of 747 amino acids. The mouse beta Mac-1 subunit is highly similar to its human counterpart with an overall sequence identity of 81% and identical positioning of 5 out of 6 potential N-linked glycosylation sites, as well as 56 Cys residues that are organized in repeating motifs characteristic of integrin beta subunits. The most highly conserved regions are the transmembrane and cytoplasmic domains where only 4 out of 69 amino acids differ, indicating that the functions associated with this domain in Mac-1-mediated processes, such as iC3b-triggered phagocytosis, have been evolutionarily conserved.