Disruption of pre-mRNA splicing in vivo results in reorganization of splicing factors.
Disruption of pre-mRNA splicing in vivo results in reorganization of splicing factors.
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DOI：
10.1083/jcb.124.3.249
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发表时间：
1994-02
期刊：
影响因子：
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通讯作者：
Spector DL
中科院分区：
文献类型：
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作者：
O'Keefe RT;Mayeda A;Sadowski CL;Krainer AR;Spector DL
我们已经研究了在哺乳动物细胞核中斑点分布的前mRNA剪接因子的组织的功能意义。在体外向活细胞显微注射抑制前mRNA剪接的寡核苷酸或抗体后，我们观察到体内剪接因子的组织发生了重大变化。免疫荧光检测显示，染色质间颗粒簇形状均匀，数量减少，剪接因子的大小和含量均增加。这些变化是短暂的，剪接因子的组织在微量注射后24小时恢复到正常分布。微量注射这些寡核苷酸或抗体也会导致体内转录的减少，但这些寡核苷酸在体外并不抑制转录。对照寡核苷酸不干扰体内的剪接或转录。我们认为，我们观察到的剪接因子的重组是体内剪接抑制的结果。
We have examined the functional significance of the organization of pre- mRNA splicing factors in a speckled distribution in the mammalian cell nucleus. Upon microinjection into living cells of oligonucleotides or antibodies that inhibit pre-mRNA splicing in vitro, we observed major changes in the organization of splicing factors in vivo. Interchromatin granule clusters became uniform in shape, decreased in number, and increased in both size and content of splicing factors, as measured by immunofluorescence. These changes were transient and the organization of splicing factors returned to their normal distribution by 24 h following microinjection. Microinjection of these oligonucleotides or antibodies also resulted in a reduction of transcription in vivo, but the oligonucleotides did not inhibit transcription in vitro. Control oligonucleotides did not disrupt splicing or transcription in vivo. We propose that the reorganization of splicing factors we observed is the result of the inhibition of splicing in vivo.