Histographic recording of human immunodeficiency virus type 1 (HIV-1) regulatory protein Rev and nuclear factors
Histographic recording of human immunodeficiency virus type 1 (HIV-1) regulatory protein Rev and nuclear factors
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DOI：
10.1007/s007050050286
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发表时间：
1998-01-01
影响因子：
2.7
通讯作者：
Haukenes, G
中科院分区：
文献类型：
--
作者：
Kaneström, A;Andresen, V;Haukenes, G
HeLa细胞和表达HIV-1调节蛋白Rev的HeLa细胞进行Rev和前mRNA加工因子的免疫染色，并通过共聚焦激光扫描显微镜进行组织学检查。在用三磷酸溴尿苷短脉冲标记新生RNA后，得到颗粒状核质染色，该染色向外周略微增加，异质性核糖核蛋白（hnRNP）A1和特别是C1/C2也是如此，其分布模式尚未描述。小核糖核蛋白颗粒（snRNP）蛋白U1、U2、U4/U6和U 5的sm-抗原除了与非snRNP剪接因子SC-35的斑点共定位的斑点外，还弥漫性地染色核质。溴化RNA和hnRNP A1和C1/C2在不同程度上被排除在斑点之外。Rev主要集中在核仁内，常呈核仁周环/带。Rev还对核质和细胞质进行染色，而不与上述蛋白质或溴化RNA共定位，并且在SC-35斑点中没有富集或排除。核仁蛋白B23和C23，像Rev，主要给了一个核周环和染色的核质，但没有以其他方式共定位与Rev或与核蛋白。组织学记录的免疫荧光图像被证明是一个有价值的工具，在本地化的HIV-1 Rev和细胞成分的研究和可能的共同定位。前mRNA加工因子与主要核仁抗原的亚细胞模式的平行比较是新的，并表明这些因子在核质中没有严格分离。
HeLa cells and HeLa cells expressing the HIV-1 regulatory protein Rev were immunostained for Rev and pre-mRNA processing factors and examined histographically by confocal laser scanning microscopy. Following short pulse-labelling with bromouridine tri-phosphate nascent RNA gave a granular nucleoplasmic staining increasing somewhat towards the periphery as did also the heterogeneous ribonucleoproteins (hnRNPs) A1 and particularly C1/C2, a distribution pattern which has not been described. The sm-antigen of the small ribonucleoprotein particle (snRNP) proteins U1, U2, U4/U6 and U5 stained the nucleoplasm diffusely in addition to speckles which co-localised with speckles of the non-snRNP splicing factor SC-35. Brominated RNA and the hnRNPs A1 and C1/C2 were to varying degrees excluded from the speckles. Rev concentrated in the nucleolus and often as a perinucleolar ring/zone. Rev also stained the nucleoplasm and cytoplasm without co-localising with the above-mentioned proteins or brominated RNA and was not enriched or excluded in SC-35 speckles. The nucleolar proteins B23 and C23, like Rev, gave primarily a perinucleolar ring and stained the nucleoplasm but did not otherwise co-localise with Rev or with nuclear proteins. Histographic recording of immunofluorescence images proved to be a valuable tool in the study of localisation of HIV-1 Rev and cellular components and of possible co-localisations. A parallel comparison of the subcellular patterns of pre-mRNA processing factors versus major nucleolar antigens is new and suggests that the factors are not strictly separated in the nucleoplasm.