A Targeted Deletion of a Region Upstream from the Jκ Cluster Impairs κ Chain Rearrangement In Cis in Mice and in the 103/bcl2 Cell Line

We have shown previously that a mutation of the KI-KII site immediately 5′ to Jκ1 on the mouse immunoglobulin light chain κ locus reduces the rearrangement level in cis, although it does not affect transcription. Here we deleted by homologous recombination in mouse embryonic stem cells a 4-kb DNA fragment, located immediately upstream of the KI-KII element, which contains the promoter of the long germline transcript. Analysis of gene-targeted heterozygous mouse splenic B cells showed a strong decrease in rearrangement for the allele bearing the deletion. When both the KI-KII mutation and the 4-kb deletion were present on the same allele, the overall reduction in rearrangement was stronger than with the 4-kb deletion alone underlying the role of these two elements in the regulation of rearrangement. The same deletion was performed by homologous recombination on one allele of the rearrangement- inducible mouse 103/bcl2-hygroR pre-B cell line, and resulted in a similar reduction in the induction of rearrangement of the mutated allele. This result validates this cell line as an in vitro model for studying the incidence of gene-targeted modifications of the κ locus on the regulation of rearrangement.

[1]  L. Ferradini,et al.  Negative regulation of Ig gene rearrangement by a 150‐bp transcriptional silencer , 1998, European journal of immunology.

[2]  A. Chess,et al.  Kappa chain monoallelic demethylation and the establishment of allelic exclusion. , 1998, Genes & development.

[3]  S. Chattopadhyay,et al.  Biochemical and functional analyses of chromatin changes at the TCR-beta gene locus during CD4-CD8- to CD4+CD8+ thymocyte differentiation. , 1998, Journal of immunology.

[4]  D. Cado,et al.  Developmentally programmed rearrangement of T cell receptor Vgamma genes is controlled by sequences immediately upstream of the Vgamma genes. , 1998, Immunity.

[5]  M. Schlissel,et al.  Accessibility and the developmental regulation of V(D)J recombination. , 1997, Seminars in immunology.

[6]  S. Takeshita,et al.  Pax‐5 is identical to EBB‐1/KLP and binds to the VpreB and λ5 promoters as well as the KI and KII sites upstream of the Jχ genes , 1997 .

[7]  F. Nato,et al.  B lineage-restricted rearrangement of a human Ig kappa transgene. , 1997, European Journal of Immunology.

[8]  B. Wasylyk,et al.  Pax-5 (BSAP) recruits Ets proto-oncogene family proteins to form functional ternary complexes on a B-cell-specific promoter. , 1996, Genes & development.

[9]  M. Singh,et al.  B cell lineage-specific activator protein (BSAP). A player at multiple stages of B cell development. , 1996, Journal of immunology.

[10]  K. Rajewsky,et al.  Rearrangement-Enhancing Element Upstream of the Mouse Immunoglobulin Kappa Chain J Cluster , 1996, Science.

[11]  I. Villey,et al.  Defect in rearrangement of the most 5' TCR-J alpha following targeted deletion of T early alpha (TEA): implications for TCR alpha locus accessibility. , 1996, Immunity.

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

[13]  D. Ramsden,et al.  Formation and resolution of double-strand break intermediates in V(D)J rearrangement. , 1995, Genes & development.

[14]  D. Loh,et al.  V(D)J recombination and allelic exclusion of a TCR beta-chain minilocus occurs in the absence of a functional promoter. , 1995, Journal of immunology.

[15]  P. Ferrier,et al.  The V(D)J recombinational and transcriptional activities of the immunoglobulin heavy-chain intronic enhancer can be mediated through distinct protein-binding sites in a transgenic substrate , 1995, Molecular and cellular biology.

[16]  L. Madisen,et al.  Identification of a locus control region in the immunoglobulin heavy-chain locus that deregulates c-myc expression in plasmacytoma and Burkitt's lymphoma cells. , 1994, Genes & development.

[17]  A. Okada,et al.  Differential activation of transcription versus recombination of transgenic T cell receptor beta variable region gene segments in B and T lineage cells , 1994, The Journal of experimental medicine.

[18]  N. Rosenberg,et al.  An active v-abl protein tyrosine kinase blocks immunoglobulin light-chain gene rearrangement. , 1994, Genes & development.

[19]  P. Tucker,et al.  Functional analysis of the V gamma 3 promoter of the murine gamma delta T-cell receptor , 1994, Molecular and cellular biology.

[20]  I. Mårtensson,et al.  Promoter, enhancer and silencer elements regulate rearrangement of an immunoglobulin transgene. , 1993, The EMBO journal.

[21]  C. Babinet,et al.  The intronic immunoglobulin ϰ gene enhancer acts independently on rearrangement and on transcription , 1993 .

[22]  F. Alt,et al.  Gene rearrangement and B-cell development. , 1993, Current opinion in immunology.

[23]  B. V. Van Ness,et al.  Initiation and processing of two kappa immunoglobulin germ line transcripts in mouse B cells , 1990, Molecular and cellular biology.

[24]  D. Baltimore,et al.  Activation of immunoglobulin kappa gene rearrangement correlates with induction of germline kappa gene transcription , 1989, Cell.

[25]  Mario R. Capecchi,et al.  Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes , 1988, Nature.

[26]  D. Baltimore,et al.  B lymphocyte-specific protein binding near an immunoglobulin kappa-chain gene J segment. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[27]  G. Krämmer,et al.  Construction of a fusion gene that confers resistance against hygromycin B to mammalian cells in culture. , 1985, Experimental cell research.

[28]  R. Perry,et al.  Transcription of the unrearranged mouse C kappa locus: sequence of the initiation region and comparison of activity with a rearranged V kappa-C kappa gene. , 1981, Cell.