首页> 美国卫生研究院文献>Molecular and Cellular Biology >Identification of two RNA-binding proteins in Balbiani ring premessenger ribonucleoprotein granules and presence of these proteins in specific subsets of heterogeneous nuclear ribonucleoprotein particles.
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Identification of two RNA-binding proteins in Balbiani ring premessenger ribonucleoprotein granules and presence of these proteins in specific subsets of heterogeneous nuclear ribonucleoprotein particles.

机译:在Balbiani环信使核糖核蛋白颗粒中鉴定出两种RNA结合蛋白并且这些蛋白在异质核糖核蛋白颗粒的特定子集中存在。

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摘要

Balbiani ring (BR) granules are premessenger ribonucleoprotein particles (RNPs) generated in giant chromosomal puffs, the BRs, in the larval salivary glands of the dipteran chironomus tentans. Monoclonal antibodies were raised against nuclear proteins collected on a single-stranded-DNA-agarose affinity column, and two of them were used to identify RNA-binding proteins in BR granules. First, in Western blots (immunoblots), one of the antibodies recognized a 36-kDa protein and the other recognized a 45-KDa protein. Second, both antibodies bound to the BRs in immunocytological experiments. It was shown in cross-linking experiments that the two proteins are associated with heterogeneous nuclear RNP (hnRNP) complexes extracted from C. tentans nuclei. By immunoelectron microscopy of isolated and partly unfolded BR RNPs, it was specifically demonstrated that the BR granules contain the two proteins and, in addition, that both proteins are distributed frequently along the RNP fiber of the particles. Thus, the 36- and 45-KDa proteins are likely to be abundant, RNA-binding proteins in the BR particles. To elucidate to what extent the two proteins are also present in other hnRNPs, we studied the binding of the antibodies to chromosomal puffs in general. It was observed that many puffs in addition to the BRs harbor the two proteins, but there are also puffs containing only one of the components, either the 36- or the 45-kDa protein. We conclude that the two proteins are not randomly bound to all hnRNPs but that each of them seems to be linked to a specific subset of the particles.
机译:Balbiani环(BR)颗粒是在二倍体chironomus tentans幼虫唾液腺中的巨大染色体泡芙(BR)中产生的信使核糖核蛋白颗粒(RNP)。产生了针对在单链DNA-琼脂糖亲和柱上收集的核蛋白的单克隆抗体,并且其中两个用于鉴定BR颗粒中的RNA结合蛋白。首先,在蛋白质印迹(免疫印迹)中,一种抗体识别36 kDa的蛋白质,另一种识别45 KDa的蛋白质。第二,在免疫细胞学实验中,两种抗体均与BR结合。在交联实验中表明,这两种蛋白质与从帐篷状衣原体核中提取的异质核RNP(hnRNP)复合物有关。通过分离的和部分未折叠的BR RNP的免疫电子显微术,特别证明了BR颗粒含有两种蛋白质,此外,这两种蛋白质经常沿颗粒的RNP纤维分布。因此,在BR颗粒中36-和45-KDa蛋白可能是丰富的RNA结合蛋白。为了阐明这两种蛋白质在其他hnRNP中也存在的程度,我们通常研究了抗体与染色体粉扑的结合。观察到,除了BR之外,还有许多粉扑还带有这两种蛋白质,但也有一些粉扑仅包含一种成分,即36 kDa或45 kDa蛋白。我们得出的结论是,这两种蛋白并不是随机地与所有hnRNP结合,但是它们每个似乎都与特定的颗粒子集相连。

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