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首页> 美国卫生研究院文献>Immunology >Germ-line human epsilon heavy chain gene RNA transcripts utilize the full range of alternative 3 splicing seen in productive epsilon mRNA.
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Germ-line human epsilon heavy chain gene RNA transcripts utilize the full range of alternative 3 splicing seen in productive epsilon mRNA.

机译：胚系人类epsilon重链基因RNA转录本利用了在生产epsilon mRNA中所见的3剪接的全部范围。

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An early step in immunoglobulin isotype switching involves the initiation of active transcription of downstream heavy chain loci while they are still in germ-line configuration. This results in the production of 'sterile' germ-line RNA transcripts that do not contain VDJ sequences, the role of which in isotype switching is under intense scrutiny. We investigated whether such human epsilon germ-line transcripts employ the full complement of complex alternative 3' RNA splicing, splicing we have recently reported occurring with productive epsilon mRNA transcripts. Using a reverse transcriptase polymerase chain reaction (RT-PCR) strategy, in which the 5' primer was located in the I region of the epsilon gene, a region expressed in germ-line but not productive (VDJ containing) epsilon transcripts, we showed that the full range of alternative 3' epsilon splices occur with germ-line transcripts. These results demonstrate that epsilon 3' splicing events are independent of 5' isotype DNA switching. Additionally, we showed that, just as with mature epsilon mRNA, the relative production of the various epsilon germ-line mRNA isoforms was responsive to modulation by stimuli such as interleukin-10 (IL-10). Thus B cells, when stimulated to produce epsilon germ-line transcripts, generate a family of germ-line mRNA that differ not only in their initiation sites but, more importantly, also differ in their 3' sequences, sequences that could be important in regulation of the parent gene itself. Furthermore, by discontinuous or trans-splicing, cells could utilize these various epsilon germ-line transcripts to produce the full range of mature IgE proteins prior to undergoing deletional recombination of isotype switching.
机译：免疫球蛋白同种型转换的早期步骤包括下游重链基因座仍处于种系构型时开始主动转录。这导致产生不包含VDJ序列的“无菌”种系RNA转录物，其在同种型转换中的作用受到严格审查。我们调查了此类人类ε生殖系转录本是否采用复杂的替代性3'RNA剪接的完整互补，我们最近报道该剪接与生产性εmRNA转录本有关。我们使用逆转录酶聚合酶链反应（RT-PCR）策略，其中5'引物位于epsilon基因的I区，即在种系中表达的区域，但不是高效的（包含VDJ的）epsilon转录物，完整的替代3'ε剪接与种系转录本一起出现。这些结果表明，ε3'剪接事件与5'同型DNA转换无关。此外，我们发现，与成熟的εmRNA一样，各种ε生殖系mRNA亚型的相对产生对白细胞介素10（IL-10）等刺激物的调节也有反应。因此，当B细胞受到刺激产生ε种系转录物时，会产生一系列的种系mRNA，它们不仅在起始位点上不同，而且更重要的是在3'序列上也不同，这些序列在调节中可能很重要亲本基因本身。此外，通过不连续或反式剪接，细胞可以在经历同种型转换的缺失重组之前，利用这些各种ε种系转录本来产生完整范围的成熟IgE蛋白。

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