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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
2
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pubmed:dateCreated |
1994-2-24
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pubmed:abstractText |
We have used extrachromosomal substrates carrying immunoglobulin heavy-chain S mu and S gamma 3 switch region sequences to study activation and targeting of recombination by a transcriptional enhancer element. Substrates are transiently introduced into activated primary murine B cells, in which recombination involving S-region sequences deletes a conditionally lethal marker, and recombination is measured by transformation of Escherichia coli in the second step of the assay. Previously we found that as many as 25% of replicated substrates recombined during 40-h transfection of lipopolysaccharide (LPS)-stimulated primary cells and that efficient recombination was dependent on the presence of S-region sequences as well as a transcriptional activator region in the constructs (H. Leung and N. Maizels, Proc. Natl. Acad. Sci. USA 89:4154-4158, 1992). Here we show that recombination of the switch substrates is threefold more efficient in LPS-cultured primary B cells than in the T-cell line EL4; the activities responsible for switch substrate recombination thus appear to be more abundant or more active in cells which can carry out chromosomal switch recombination. We test the role of the transcriptional activator region and show that the immunoglobulin heavy-chain intron enhancer (E mu) alone stimulates recombination as well as E mu combined with a heavy-chain promoter and that mutations that diminish enhancer-dependent transcription 500-fold diminish recombinational activation less than 2-fold. These observations suggest that the enhancer stimulates recombination by a mechanism that does not depend on transcript production or that is insensitive to the level of transcript production over a very broad range. Furthermore, we find that E mu stimulates recombination when located either upstream or downstream of S mu but that the position of the recombinational activator does affect the targeting of recombination junctions, suggesting that the relatively imprecise targeting of switch junctions in vivo may reflect the availability of many potential activator sites within each switch region.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1315054,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1349323,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1408813,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1459429,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1500850,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1505030,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1516825,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1552290,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1555239,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1639801,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1690849,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1714414,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1748289,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-1986228,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-2111440,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-2116362,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-2188677,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-2511438,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-2528697,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-2578321,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-2694945,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-2777075,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-2838451,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-2966762,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-2972385,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-2987353,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-3016509,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-3034595,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-3122021,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-3132612,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-3317825,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-6288254,
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http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-6960240,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-8253079,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-8341678,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-8355709,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-8438159,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-8441648,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8289820-8514805
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Feb
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pubmed:issn |
0270-7306
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
14
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1450-8
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:8289820-Animals,
pubmed-meshheading:8289820-B-Lymphocytes,
pubmed-meshheading:8289820-Base Sequence,
pubmed-meshheading:8289820-Cell Line,
pubmed-meshheading:8289820-Cells, Cultured,
pubmed-meshheading:8289820-DNA Primers,
pubmed-meshheading:8289820-Enhancer Elements, Genetic,
pubmed-meshheading:8289820-Gene Expression Regulation,
pubmed-meshheading:8289820-Immunoglobulin Heavy Chains,
pubmed-meshheading:8289820-Immunoglobulin Switch Region,
pubmed-meshheading:8289820-Immunoglobulin gamma-Chains,
pubmed-meshheading:8289820-Immunoglobulin mu-Chains,
pubmed-meshheading:8289820-Mice,
pubmed-meshheading:8289820-Molecular Sequence Data,
pubmed-meshheading:8289820-Mutagenesis, Insertional,
pubmed-meshheading:8289820-Plasmids,
pubmed-meshheading:8289820-Promoter Regions, Genetic,
pubmed-meshheading:8289820-Recombination, Genetic,
pubmed-meshheading:8289820-Restriction Mapping,
pubmed-meshheading:8289820-T-Lymphocytes,
pubmed-meshheading:8289820-Thymoma,
pubmed-meshheading:8289820-Thymus Neoplasms,
pubmed-meshheading:8289820-Transfection,
pubmed-meshheading:8289820-Tumor Cells, Cultured
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pubmed:year |
1994
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pubmed:articleTitle |
Regulation and targeting of recombination in extrachromosomal substrates carrying immunoglobulin switch region sequences.
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pubmed:affiliation |
Department of Molecular Biophysics and Biochemistry, Yale Medical School, New Haven, Connecticut 06510.
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pubmed:publicationType |
Journal Article,
Comparative Study
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