Different CD40-mediated signaling events require distinct CD40 structural features.
Signals delivered to B cells through CD40 are critical to the development of humoral immune responses. In this study we characterize regions of the 62-amino acid cytoplasmic domain of human CD40 (hCD40) that are essential for signal transduction and examine the functional consequences of mutations in these regions. A panel of mutant hCD40 molecules was stably expressed in mouse B cell lines and tested for its ability to stimulate Ab secretion, homotypic adhesion, and increased surface expression of B7, Fas, CD23, LFA-1, and intracellular adhesion molecule-1. Our results indicate that CD40 contains at least two major signaling determinants in the cytoplasmic domain: one disrupted by a truncation of 22 amino acids, and a second disrupted by the removal of 10 additional amino acids. The second determinant includes threonine 234, a residue previously shown to be important in CD40 signal transduction. Our functional analysis of alanine and serine substitutions at position 234 indicates that phosphorylation of this residue may be important for full CD40 signaling activity.
Hostager
B S
BS
Department of Microbiology, University of Iowa, Iowa City, IA 52242, USA.
Hsing
Y
Y
Harms
D E
DE
Bishop
G A
GA
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D019013:CD40 Antigens / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D002448:Cell Adhesion; D002449:Cell Aggregation; D002460:Cell Line; D006224:Cricetinae; D005434:Flow Cytometry; D006801:Humans; D007075:Immunoglobulin M / Q000378:metabolism; D018799:Intercellular Adhesion Molecule-1; D016169:Lymphocyte Function-Associated Antigen-1 / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017455:Receptors, IgE / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D013329:Structure-Activity Relationship; D019014:fas Receptor / Q000378:metabolism
D019013:CD40 Antigens; D007075:Immunoglobulin M; D016169:Lymphocyte Function-Associated Antigen-1; D017455:Receptors, IgE; D019014:fas Receptor; D018799:Intercellular Adhesion Molecule-1