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首页> 外文期刊>Nucleic Acids Research >V(D)J RECOMBINATION FREQUENCY IS AFFECTED BY THE SEQUENCE INTERPOSED BETWEEN A PAIR OF RECOMBINATION SIGNALS - SEQUENCE COMPARISON REVEALS A PUTATIVE RECOMBINATIONAL ENHANCER ELEMENT
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V(D)J RECOMBINATION FREQUENCY IS AFFECTED BY THE SEQUENCE INTERPOSED BETWEEN A PAIR OF RECOMBINATION SIGNALS - SEQUENCE COMPARISON REVEALS A PUTATIVE RECOMBINATIONAL ENHANCER ELEMENT

机译：V（D）J重组频率受到一对重组信号之间插入的序列的影响-序列比较揭示了一种有效的重组增强元件

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The immunoglobulin heavy chain intron enhancer (E mu) not only stimulates transcription but also V(D)J recombination of chromosomally integrated recombination substrates. We aimed at reproducing this effect in recombination competent cells by transient transfection of extrachromosomal substrates. These we prepared by interposing between the recombination signal sequences (RSS) of the plasmid pBlueRec various fragments, including E mu, possibly affecting V(D)J recombination, Our work shows that sequences inserted between RSS 23 and RSS 12, with distances from their proximal ends of 26 and 284 bp respectively, can markedly affect the frequency of V(D)J recombination. We report that the entire E mu, the E mu core as well as its flanking 5' and 3' matrix associated regions (5' and 3' MARs) upregulate V(D)J recombination while the downstream section of the 3' MAR of E mu does not. Also, prokaryotic sequences markedly suppress V(D)J recombination. This confirms previous results obtained with chromosomally integrated substrates, except for the finding that the full length 3' MAR of E mu stimulates V(D)J recombination in an episomal but not in a chromosomal context. The fact that other MARs do not share this activity suggests that the effect is not mediated through attachment of the recombination substrate to a nuclear matrix-associated recombination complex but through cis-activation. The presence of a 26 bp A-T-rich sequence motif in the 5' and 3' MARs of E mu and in all of the other upregulating fragments investigated, leads us to propose that the motif represents a novel recombinational enhancer element distinct from those constituting the E mu core.
机译：免疫球蛋白重链内含子增强子（E mu）不仅刺激转录，而且刺激染色体整合重组底物的V（D）J重组。我们旨在通过染色体外底物的瞬时转染在重组感受态细胞中重现这一效应。这些是通过在质粒pBlueRec的重组信号序列（RSS）之间插入各种片段（包括Emu，可能影响V（D）J重组）而制备的。我们的工作表明，插入RSS 23和RSS 12之间的序列与它们之间的距离近端分别为26和284 bp，可显着影响V（D）J重组的频率。我们报告说，整个Emu，Emu核心及其侧翼的5'和3'矩阵相关区域（5'和3'MARs）上调V（D）J重组，而3'MAR的下游部分mu没有。此外，原核序列显着抑制V（D）J重组。这证实了用染色体整合的底物获得的先前结果，除了发现E mu的全长3'MAR刺激了游离型而不是染色体上的V（D）J重组。其他MAR不共享此活性的事实表明，该作用不是通过重组底物与核基质相关重组复合物的附着介导，而是通过顺式激活。 E mu的5'和3'MAR中以及所有其他上调片段中均存在26 bp富AT序列的基序，这使我们提出，该基序代表了一种新颖的重组增强子元件，与构成该蛋白的重组增强子不同。 mu亩芯。

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