Immunoglobulin mu heavy chains do not mediate tyrosine phosphorylation of Ig alpha from the ER-cis-Golgi.
Signals delivered by Ig receptors guide the development of functional B lymphocytes. For example, clonal expansion of early mu heavy chain ( mu HC)-positive pre-B cells requires the assembly of a signal-competent pre-B cell receptor complex (pre-BCR) consisting of a mu HC, a surrogate L chain, and the signal dimer Ig alpha beta. However, only a small fraction of the pre-BCR is transported to the cell surface, suggesting that pre-BCR signaling initiates already from an intracellular compartment, e.g., the endoplasmic reticulum (ER). The finding that differentiation of pre-B cells and allelic exclusion at the IgH locus take place in surrogate L chain-deficient mice further supports the presence of a mu HC-mediated intracellular signal pathway. To determine whether a signal-competent Ig complex can already be assembled in the ER, we analyzed the consequence of pervanadate on tyrosine phosphorylation of Ig alpha in J558L plasmacytoma and 38B9 pre-B cells transfected with either a transport-competent IgL chain-pairing or an ER-retained nonpairing micro HC. Flow cytometry, combined Western blot-immunoprecipitation-kinase assays, and confocal microscopy revealed that both the nonpairing and pairing mu HC assembled with the Ig alpha beta dimer; however, in contrast to a pairing mu HC, the nonpairing mu HC was retained in the ER-cis-Golgi compartment, and neither colocalized with the src kinase lyn nor induced tyrosine phosphorylation of Ig alpha after pervanadate treatment of cells. On the basis of these findings, we propose that a signal-competent Ig complex consisting of mu HC, Ig alpha beta, and associated kinases is assembled in a post-ER compartment, thereby supporting the idea that a pre-BCR must be transported to the cell surface to initiate pre-BCR signaling.
Mielenz
Dirk
D
Division of Molecular Immunology, Department of Internal Medicine III, Nikolaus-Fiebiger Center, University of Erlangen-Nürnberg, Erlangen, Germany.
Ruschel
Anja
A
Vettermann
Christian
C
Jäck
Hans-Martin
HM
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D016175:B-Lymphocyte Subsets / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D051925:CD79 Antigens; D045744:Cell Line, Tumor; D002999:Clone Cells; D003593:Cytoplasm / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D004789:Enzyme Activation / Q000276:immunology; D006056:Golgi Apparatus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D007148:Immunoglobulin mu-Chains / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D051379:Mice; D010766:Phosphorylation / Q000187:drug effects; D021381:Protein Transport / Q000235:genetics / Q000276:immunology; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism*; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology; D013234:Stem Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology; D014443:Tyrosine / Q000378:metabolism*; D014638:Vanadates / Q000494:pharmacology; D019061:src-Family Kinases / Q000378:metabolism
D015703:Antigens, CD; D051925:CD79 Antigens; C497882:Cd79a protein, mouse; C497884:Cd79b protein, mouse; D007143:Immunoglobulin Heavy Chains; D007148:Immunoglobulin mu-Chains; D011947:Receptors, Antigen, B-Cell; C076715:pervanadate; D014638:Vanadates; D014443:Tyrosine; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases