INVIVO DETECTION OF SNRNP-RICH ORGANELLES IN THE NUCLEI OF MAMMALIAN-CELLS
INVIVO DETECTION OF SNRNP-RICH ORGANELLES IN THE NUCLEI OF MAMMALIAN-CELLS
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DOI：
10.1002/j.1460-2075.1991.tb07712.x
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发表时间：
1991-07-01
期刊：
影响因子：
11.4
通讯作者：
LAMOND, AI
中科院分区：
文献类型：
--
作者：
CARMOFONSECA, M;PEPPERKOK, R;LAMOND, AI
通过将荧光标记的反义探针微注射到活HeLa和3T3细胞的细胞核中，分析了snRNPs在体内的分布。U2和U5 snRNA的探针特异性标记相同的离散核病灶，而U1 snRNA的探针除了标记病灶外，还显示广泛的核质标记，不包括核核。U3 snRNA探针特异标记核仁。这些体内数据证实，哺乳动物细胞具有含有剪接体snRNPs的核灶。体内和原位共定位研究表明，剪接体snrna存在于相同的核病灶中。这些病灶也被识别snRNP蛋白、m3G-cap结构和剪接因子U2AF的抗体染色，但不被抗sc -35或抗la抗体染色。在放线菌素D治疗前后，U1 snRNP和剪接因子U2AF在细胞核中紧密共定位，表明它们在体内可能是同一个复合体的一部分。
The in vivo distribution of snRNPs has been analysed by microinjecting fluorochrome-labelled antisense probes into the nuclei of live HeLa and 3T3 cells. Probes for U2 and U5 snRNAs specifically label the same discrete nuclear foci while a probe for U1 snRNA shows widespread nucleoplasmic labelling, excluding nucleoli, in addition to labelling foci. A probe for U3 snRNA specifically labels nucleoli. These in vivo data confirm that mammalian cells have nuclear foci which contain spliceosomal snRNPs. Co-localization studies, both in vivo and in situ, demonstrate that the spliceosomal snRNAs are present in the same nuclear foci. These foci are also stained by antibodies which recognize snRNP proteins, m3G-cap structures and the splicing factor U2AF but are not stained by anti-SC-35 or anti-La antibodies. U1 snRNP and the splicing factor U2AF closely co-localize in the nucleus, both before and after actinomycin D treatment, suggesting that they may both be part of the same complex in vivo.