title: Down-regulation of RAG1 and RAG2 gene expression in preB cells after functional immunoglobulin heavy chain rearrangement.
abstract: Two waves of immunoglobulin gene rearrangements, first of the heavy, then of the light chain chain gene loci form functional immunoglobulin genes during B cell development. In mouse bone marrow the differential surface expression of B220 (CD45R), c-kit, CD25, and surrogate light chain as well as the cell cycle status allows FACS separation of the cells in which these two waves of rearrangements occur. The gene products of two recombination activating genes, RAG1 and RAG2 are crucial for this rearrangement process. Here, we show that the expression of the RAG genes is twice up- and down-regulated, at the transcriptional level for RAG1 and RAG2, and at the postranscriptional level for RAG2 protein. Expression levels are high in D-->JH and VH-->DJH rearranging proB and preB-I cells, low in preB cells expressing the preB cell receptor on the cell surface, and high again in VL-->JL rearranging small preB-II cells. In immature B cells expressing on the cell surface RAG1 and RAG2 mRNA is down-regulated, whereas RAG2 protein levels are maintained. Down-regulation of RAG1 and RAG2 gene expression after productive rearrangement at one heavy chain allele might be part of the mechanisms that prevent further rearrangements at the other allele.
authors.affiliation: Basel Institute for Immunology, Switzerland.
authors.forename: U
authors.initials: U
authors.lastname: Grawunder
authors.forename: T M
authors.initials: TM
authors.lastname: Leu
authors.forename: D G
authors.initials: DG
authors.lastname: Schatz
authors.forename: A
authors.initials: A
authors.lastname: Werner
authors.forename: A G
authors.initials: AG
authors.lastname: Rolink
authors.forename: F
authors.initials: F
authors.lastname: Melchers
authors.forename: T H
authors.initials: TH
authors.lastname: Winkler
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000502:physiology; D001483:Base Sequence; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D004268:DNA-Binding Proteins; D015536:Down-Regulation / Q000235:genetics*; D005260:Female; D005434:Flow Cytometry; D015322:Gene Rearrangement, B-Lymphocyte / Q000235:genetics*; D018398:Homeodomain Proteins; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics; D019009:Proto-Oncogene Proteins c-kit / Q000502:physiology; D012333:RNA, Messenger / Q000032:analysis; D011947:Receptors, Antigen, B-Cell / Q000096:biosynthesis
chemical_list: D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D007143:Immunoglobulin Heavy Chains; D011506:Proteins; D012333:RNA, Messenger; C499308:Rag2 protein, mouse; D011947:Receptors, Antigen, B-Cell; C064655:V(D)J recombination activating protein 2; C064658:RAG-1 protein; D019009:Proto-Oncogene Proteins c-kit