Localized Gene-Specific Induction of Accessibility to V(D)j Recombination Induced by E2a and Early B Cell Factor in Nonlymphoid Cells

Accessibility of immunoglobulin (Ig) gene segments to V(D)J recombination is highly regulated and is normally only achieved in B cell precursors. We previously showed that ectopic expression of E2A or early B cell factor (EBF) with recombination activating gene (RAG) induces rearrangement of IgH and IgL genes in nonlymphoid cells. VκI genes throughout the locus were induced to rearrange after transfection with E2A, suggesting that the entire Vκ locus was accessible. However, here we show that Ig loci are not opened globally but that recombination is localized. Gene families are interspersed in the DH, Vκ, and Vλ loci, and we show that certain families and individual genes undergo high levels of recombination after ectopic expression of E2A or EBF, while other families within the same locus are not induced to rearrange. Furthermore, in some families, induction of germline transcription correlates with the level of induced recombination, while in others there is no correlation, suggesting that recombination is not simply initiated by induction of germline transcription. The induced repertoire seen at 24 hours does not change significantly over time indicating the absence of many secondary rearrangements and also suggesting a direct targeting mechanism. We propose that accessibility occurs in a local manner, and that binding sites for factors facilitating accessibility are therefore likely to be associated with individual gene segments.

[1]  C. Murre,et al.  Induction of a Diverse T Cell Receptor γ/δ Repertoire by the Helix-Loop-Helix Proteins E2a and Heb in Nonlymphoid Cells , 2001, The Journal of experimental medicine.

[2]  U. Storb,et al.  Insertion of Phosphoglycerine Kinase (Pgk)-Neo 5′ of Jλ1 Dramatically Enhances Vjλ1 Rearrangement , 2001, The Journal of experimental medicine.

[3]  A. Feeney,et al.  B‐cell repertoire formation: role of the recombination signal sequence in non‐random V segment utilization , 2000, Immunological reviews.

[4]  A. Feeney,et al.  E2A and EBF act in synergy with the V(D)J recombinase to generate a diverse immunoglobulin repertoire in nonlymphoid cells. , 2000, Molecular cell.

[5]  M. Krangel,et al.  A role for histone acetylation in the developmental regulation of VDJ recombination. , 2000, Science.

[6]  S. Berger,et al.  A conserved motif present in a class of helix-loop-helix proteins activates transcription by direct recruitment of the SAGA complex. , 1999, Molecular cell.

[7]  J. V. van Dongen,et al.  Ig heavy chain gene rearrangements in T-cell acute lymphoblastic leukemia exhibit predominant DH6-19 and DH7-27 gene usage, can result in complete V-D-J rearrangements, and are rare in T-cell receptor alpha beta lineage. , 1999, Blood.

[8]  P. Lipsky,et al.  Molecular Mechanisms and Selection Influence the Generation of the Human VλJλ Repertoire , 1999, The Journal of Immunology.

[9]  C. Murre,et al.  Positive and Negative Regulation of  V(D)J Recombination by the E2A Proteins , 1999, The Journal of experimental medicine.

[10]  K. Calame,et al.  Evidence that Immunoglobulin VH-DJ Recombination Does Not Require Germ Line Transcription of the Recombining Variable Gene Segment , 1998, Molecular and Cellular Biology.

[11]  A. Okada,et al.  Accessibility control of variable region gene assembly during T‐cell development , 1998, Immunological reviews.

[12]  C. Murre,et al.  Induction of Early B Cell Factor (EBF) and Multiple B Lineage Genes by the Basic Helix-Loop-Helix Transcription Factor E12 , 1998, The Journal of experimental medicine.

[13]  D. Cado,et al.  Developmentally Programmed Rearrangement of T Cell Receptor Vγ Genes Is Controlled by Sequences Immediately Upstream of the Vγ Genes , 1998 .

[14]  B. Nadel,et al.  Sequence of the Spacer in the Recombination Signal Sequence Affects V(D)J Rearrangement Frequency and Correlates with Nonrandom Vκ Usage In Vivo , 1998, The Journal of experimental medicine.

[15]  A. Riddell,et al.  Impaired immunoglobulin gene rearrangement in mice lacking the IL-7 receptor , 1998, Nature.

[16]  E L Sonnhammer,et al.  Sequence of the human immunoglobulin diversity (D) segment locus: a systematic analysis provides no evidence for the use of DIR segments, inverted D segments, "minor" D segments or D-D recombination. , 1997, Journal of molecular biology.

[17]  D. Baltimore,et al.  Complementation of V(D)J recombination deficiency in RAG-1(-/-) B cells reveals a requirement for novel elements in the N-terminus of RAG-1. , 1997, Immunity.

[18]  R. Brezinschek,et al.  Analysis of the human VH gene repertoire. Differential effects of selection and somatic hypermutation on human peripheral CD5(+)/IgM+ and CD5(-)/IgM+ B cells. , 1997, The Journal of clinical investigation.

[19]  I. Villey,et al.  Defect in rearrangement of the most 5′TCR-jα following targeted deletion of T early α (tea): Implications for TCRα locus accessibility , 1997 .

[20]  P. T. Jones,et al.  The creation of diversity in the human immunoglobulin Vλ repertoire , 1997 .

[21]  A. Feeney,et al.  Human cord blood kappa repertoire. , 1997, Journal of immunology.

[22]  R. Brezinschek,et al.  Molecular mechanisms and selective influences that shape the kappa gene repertoire of IgM+ B cells. , 1997, The Journal of clinical investigation.

[23]  S Minoshima,et al.  One-megabase sequence analysis of the human immunoglobulin lambda gene locus. , 1997, Genome research.

[24]  M. Schlissel,et al.  Changes in Locus-specific V(D)J Recombinase Activity Induced by Immunoglobulin Gene Products during B Cell Development , 1997, The Journal of experimental medicine.

[25]  I. Villey,et al.  Defect in Rearrangement of the Most 5′ TCR–Jα Following Targeted Deletion of T Early α (TEA): Implications for TCR α Locus Accessibility , 1996 .

[26]  A. Shaffer,et al.  Cell Type–Specific Chromatin Structure Determines the Targeting of V(D)J Recombinase Activity In Vitro , 1996, Cell.

[27]  F. Alt,et al.  Deletion of the Igκ Light Chain Intronic Enhancer/Matrix Attachment Region Impairs but Does Not Abolish VκJκ Rearrangement , 1996 .

[28]  A. Marshall,et al.  Frequency of VH81x usage during B cell development: initial decline in usage is independent of Ig heavy chain cell surface expression. , 1996, Journal of immunology.

[29]  R. Grosschedl,et al.  Failure of B-cell differentiation in mice lacking the transcription factor EBF , 1995, Nature.

[30]  N. Berinstein,et al.  A mutated promoter of a human Ig V lambda gene segment is associated with reduced germ-line transcription and a low frequency of rearrangement. , 1995, Journal of immunology.

[31]  Ian Krop,et al.  E2A proteins are required for proper B cell development and initiation of immunoglobulin gene rearrangements , 1994, Cell.

[32]  H. Weintraub,et al.  The helix-loop-helix gene E2A is required for B cell formation , 1994, Cell.

[33]  H. Zachau,et al.  The variable genes of the human immunoglobulin kappa locus. , 1993, Biological chemistry Hoppe-Seyler.

[34]  M. Madaio,et al.  Molecular analysis of spontaneous nephrotropic anti-laminin antibodies in an autoimmune MRL-lpr/lpr mouse. , 1993, Journal of immunology.

[35]  F. Sablitzky,et al.  V(D)J recombination in B cells is impaired but not blocked by targeted deletion of the immunoglobulin heavy chain intron enhancer. , 1993, The EMBO journal.

[36]  J. Goldman,et al.  Ordered rearrangement of variable region genes of the T cell receptor gamma locus correlates with transcription of the unrearranged genes , 1993, The Journal of experimental medicine.

[37]  D. Baltimore,et al.  Helix-loop-helix transcription factor E47 activates germ-line immunoglobulin heavy-chain gene transcription and rearrangement in a pre-T-cell line. , 1991, Genes & development.

[38]  D. Baltimore,et al.  Virus-transformed pre-B cells show ordered activation but not inactivation of immunoglobulin gene rearrangement and transcription , 1991, The Journal of experimental medicine.

[39]  M. Yamada,et al.  Preferential utilization of specific immunoglobulin heavy chain diversity and joining segments in adult human peripheral blood B lymphocytes , 1991, The Journal of experimental medicine.

[40]  J. Pelkonen,et al.  First T cell receptor alpha gene rearrangements during T cell ontogeny skew to the 5' region of the J alpha locus. , 1990, Journal of immunology.

[41]  David Baltimore,et al.  A new DNA binding and dimerization motif in immunoglobulin enhancer binding, daughterless, MyoD, and myc proteins , 1989, Cell.

[42]  J. Allison,et al.  Developmentally ordered appearance of thymocytes expressing different T-cell antigen receptors , 1988, Nature.

[43]  T. Leanderson,et al.  Conserved sequence elements in K promoters from mice and humans: implications for transcriptional regulation and repertoire expression , 1998, Immunogenetics.

[44]  F. Alt,et al.  Accessibility control of antigen-receptor variable-region gene assembly: role of cis-acting elements. , 1996, Annual review of immunology.

[45]  G. Nolan,et al.  Production of high-titer helper-free retroviruses by transient transfection. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[46]  F. Alt,et al.  Regulation of the assembly and expression of variable-region genes. , 1986, Annual review of immunology.