Inducible DNA-protein interactions of the murine kappa immunoglobulin enhancer in intact cells: comparison with<i>in vitro</i>interactions

Hromas, Robert; Pauli, Urs; Marcuzzi, Adriana; Lafrenz, D; Nick, Harry S.; Stein, Janet L.; Stein, Gary S.; Ness, Brian Van

doi:10.1093/nar/16.3.953

Cited by 17 publications

(14 citation statements)

References 28 publications

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“…A footprint was also observed over an A/T-rich sequence situated between the E* and EO sites which does not contain defined factor binding sites. As previously reported, we could not detect protein interaction with the E1 and E2 boxes using this technique (32).…”

Section: The Mouse and Rabbit Enhancers Do Not Interact With The Samesupporting

confidence: 48%

Comparison of Mouse and Rabbit Eiκ Enhancers Indicates That Different Elements Within the Enhancer May Mediate Activation of Transcription and Recombination

Coquilleau

Cavelier

Rougeon

et al. 2000

The Journal of Immunology

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The intronic Ig κ-light chain enhancer (Eiκ) has been implicated in regulation of transcription and Vκ-Jκ recombination at the κ locus. To identify sequences within the Eiκ enhancer which are involved in control of recombination, we have made use of the finding that the Eiκ element from the rabbit b9 κ locus is capable of inducing rearrangement, but not transcription of κ genes in mouse lymphoid cells. We have therefore compared the binding of murine nuclear proteins to the mouse and rabbit Eiκ elements. DNase I footprinting and gel mobility shift assays indicate that only the κB, κE1, and κE2 sites of the rabbit enhancer are able to interact with murine trans-acting factors. Moreover, although the rabbit κB site binds murine NF-κB p50/p50 and p50/p65 complexes with high affinity, this site is not capable of mediating transcriptional activation of transient transfection reporter constructs in mouse B lineage cells. These results therefore suggest that, in contrast to the maintenance of κ enhancer transcription which requires all of the Eiκ sites, only the κB, κE1, and κE2 sites may be necessary for the recombinational activity of the enhancer. Furthermore, NF-κB-mediated effects on transcription and recombination appear to involve separate downstream activation pathways.

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Section: The Mouse and Rabbit Enhancers Do Not Interact With The Samesupporting

confidence: 48%

Comparison of Mouse and Rabbit Eiκ Enhancers Indicates That Different Elements Within the Enhancer May Mediate Activation of Transcription and Recombination

Coquilleau

Cavelier

Rougeon

et al. 2000

The Journal of Immunology

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“…Additional proteins which bind within this region of the kappa enhancer have been observed (12,23,24). Currie and Roeder have identified a nuclear factor, NF-KE*, which was found to be more abundant in HeLa cells than in B cells (12).…”

Section: Discussionmentioning

confidence: 99%

Silencing of the expression of the immunoglobulin kappa gene in non-B cells.

Pierce¹,

Gifford²,

Baltimore³

1991

Mol. Cell. Biol.

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Although the activating factor NF-cB can be present in the nucleus of many cell types, transcription and rearrangement of the immunoglobulin kappa chain gene is restricted to cells of the B lineage. Part of this specificity is determined by sequences within the major intron of the kappa gene that specifically silence gene expression in non-B cells (T cells and HeLa cells). These sequences are found in a 232-bp fragment located 5' of the NF-KB binding sequence of the enhancer. When this fragment is added back upstream of an active NF-KB site, it specifically decreases the expression of a linked gene by more than 10-fold in activated T cells but it has no effect on expression in B cells. The kappa silencer region acts in an orientation-and distanceindependent manner and appears to be composed of multiple negative elements. The kappa silencer may act to restrict transcription and rearrangement of the CK locus to cells of the B lineage.The immunoglobulin kappa chain gene is under precise developmental control. This gene is expressed only in cells of the B-lymphocyte lineage. The transcription of the murine kappa locus is regulated by the binding of nuclear factors to DNA sequences within the V,< promoters and the two enhancer regions, one located within the J-C,K intron and another located downstream of the CK coding region (5,13,15,32,40,42). Several regulatory sequences within the intron enhancer are important for enhancer function in B cells (14,27,46). These include the E motifs (KE1, KE2, and KE3), which are homologous to the protein-binding consensus sequence CAGGTGGC in the immunoglobulin heavychain enhancer and the kB site, GGGACTTTCC, which binds the nuclear factor NF-KB (46). The activation of both CK transcription and rearrangement occurs at the pre-B-to B-cell transition and is correlated with the induction of NF-KB (37,39,47,51 including interleukin-2 receptor alpha chain (7). Significantly, NF-KB can also activate several viral enhancers, including the human immunodeficiency virus enhancer, the simian virus 40 enhancer, and the cytomegalovirus enhancer (8,33).Since NF-KB can stimulate so many different genes in T and B cells, it is clear that NF-KB by itself cannot account for the precise tissue-specific regulation of each of these genes. There must be additional regulatory elements which modify the activity of NF-KB and prevent the inappropriate activation of all these genes at once. We and others have previously observed that the NF-KB binding sequence alone can activate transcription in either B cells or non-B cells treated with PMA (36,41,56

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“…Protein concentrations were determined as described by Lowry (44). Nuclear binding factors were analyzed by their altered mobility on acrylamide gels as previously described (34). Double stranded oligonucleotides used included a 34 bp sequence containing the NF-xB binding site (34, sense strand: 5'-TCTCAACAGAGGGGACTTTCCGAGAGCCATCTGG-3') and a 27 bp duplex containing the octamer sequence motif in the context of the 70Z/3 kappa promoter sequence (25, sense strand: 5'-GTATCTTGC-GATTTGCATATTACATTT-3').…”

Section: Northern Blot Analysismentioning

confidence: 99%

“…Within the n enhancer, one sequence motif which appears essential for enhancer function is identical to the 11 bp SV-40 core enhancer sequence, and a specific factor whose binding correlates with K expression has been identified and designated NF-nB (31)(32)(33). This factor has been demonstrated in mature B cell extracts and shown to be inducible by LPS in the pre-B cell line, 70Z/3 as well as normal B lymphocytes (34).…”

mentioning

confidence: 99%

Effects of anti-IgM suppression on polyclonally activated murine B cells: analysis of immunoglobulin mRNA, gene specific nuclear factors and cell cycle distribution

et al. 1989

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Polyclonal activation of murine B cells with bacterial lipopolysaccharide (LPS) and dextran sulfate (DxS) results in cell proliferation as well as increased immunoglobulin gene transcription and antibody secretion. When added to B cell cultures during mitogen activation, anti-mu antibody suppresses the rate of proliferation and immunoglobulin gene expression. Using this model system we show that co-cultures of B cells with LPS/DxS and anti-mu resulted in a decrease of both mu and kappa chain mRNA. Suppression did not prevent B cell entry into cycle nor a significant alteration in the distribution of cells in phases of cell cycle, although it did prolong the cycle transit time in a dose dependent fashion as determined by bromodeoxyuridine pulse labelling. Analysis of B cell specific nuclear binding factors, which previously have been shown to be important in regulating immunoglobulin gene transcription were examined. Results show that the kappa-specific enhancer binding activity of NF-kappa B was induced in activated as well as suppressed cultures. The lymphoid specific factor NF-A2, which recognizes the octamer sequence motif in the promoters of immunoglobulin genes, was induced by the polyclonal activation but was selectively lost in extracts from suppressed cells. Thus, specific regulation of the nuclear factor which plays a critical role in both heavy and light chain immunoglobulin gene expression may contribute to the transcriptional suppression observed in anti-mu treated B cells.

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Inducible DNA-protein interactions of the murine kappa immunoglobulin enhancer in intact cells: comparison within vitrointeractions

Cited by 17 publications

References 28 publications

Comparison of Mouse and Rabbit Eiκ Enhancers Indicates That Different Elements Within the Enhancer May Mediate Activation of Transcription and Recombination

Comparison of Mouse and Rabbit Eiκ Enhancers Indicates That Different Elements Within the Enhancer May Mediate Activation of Transcription and Recombination

Silencing of the expression of the immunoglobulin kappa gene in non-B cells.

Effects of anti-IgM suppression on polyclonally activated murine B cells: analysis of immunoglobulin mRNA, gene specific nuclear factors and cell cycle distribution

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