CD45 function is regulated by an acidic 19-amino acid insert in domain II that serves as a binding and phosphoacceptor site for casein kinase 2.
In this study experiments were conducted to elucidate the physical/functional relationship between CD45 and casein kinase 2 (CK2). Immunoprecipitation experiments demonstrated that CK2 associates with CD45 and that this interaction is inducible upon Ag receptor cross-linking in B and T cell lines as well as murine thymocytes and splenic B cells. However, yeast two-hybrid analysis failed to demonstrate a physical interaction between the individual CK2 alpha, alpha', or beta subunits and CD45. In contrast, a yeast three-hybrid assay in which either CK2 alpha and beta or alpha' and beta subunits were coexpressed with the cytoplasmic domain of CD45, demonstrated that both CK2 subunits are necessary for the interaction with CD45. Experiments using the yeast three-hybrid assay also revealed that a 19-aa acidic insert in domain II of CD45 mediates the physical interaction between CK2 and CD45. Structure/function experiments in which wild-type or mutant CD45RA and CD45RO isoforms were expressed in CD45-deficient Jurkat cells revealed that the 19-aa insert is important for optimal CD45 function. The ability of both CD45RA and CD45RO to reconstitute CD3-mediated signaling based on measurement of calcium mobilization and mitogen-activated protein kinase activation was significantly decreased by deletion of the 19-aa insert. Mutation of four serine residues within the 19-aa insert to alanine affected CD45 function to a similar extent compared with that of the deletion mutants. These findings support the hypothesis that a physical interaction between the CD45 cytoplasmic domain and CK2 is important for post-translational modification of CD45, which, in turn, regulates its catalytic function.
Greer
S F
SF
Department of Microbiology, Division of Developmental and Clinical Immunology, University of Alabama, Birmingham, AL 35294, USA.
Wang
Y
Y
Raman
C
C
Justement
L B
LB
D000818:Animals; D001402:B-Lymphocytes / Q000201:enzymology / Q000276:immunology; D001665:Binding Sites / Q000235:genetics / Q000276:immunology; D047390:Casein Kinase II; D020035:Holoenzymes / Q000378:metabolism / Q000502:physiology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D019169:Jurkat Cells; D017493:Leukocyte Common Antigens / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D051379:Mice; D016254:Mutagenesis, Insertional; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000235:genetics; D017027:Protein Tyrosine Phosphatases / Q000235:genetics / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000378:metabolism / Q000502:physiology; D017384:Sequence Deletion; D012694:Serine / Q000235:genetics; D013154:Spleen / Q000166:cytology / Q000201:enzymology / Q000276:immunology; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000201:enzymology / Q000276:immunology; D013950:Thymus Gland / Q000166:cytology / Q000201:enzymology / Q000276:immunology; D014407:Tumor Cells, Cultured
D020035:Holoenzymes; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D012694:Serine; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D017493:Leukocyte Common Antigens; D017027:Protein Tyrosine Phosphatases