Bonneaud N, Minvielle-Sebastia L, Cullin C, Lacroute F
Centre de Génétique Moléculaire du CNRS, Laboratoire Propre Associé à l'Université Pierre-et-Marie-Curie, Gif-sur-Yvette, France.
J Cell Sci. 1994 Apr;107 ( Pt 4):913-21. doi: 10.1242/jcs.107.4.913.
RNA14 and RNA15 were originally identified by temperature-sensitive mutations that cause a rapid decrease in poly(A)-tail length and overall mRNA levels at the restrictive temperature. We have raised antibodies to the RNA14 and RNA15 proteins, and used subcellular fractionation and immunofluorescence to localize these proteins within the yeast cell. RNA14p is a 73 kDa protein found in both the nucleus and the cytoplasm, whilst RNA15p is a 42 kDa protein detected only in the nucleus. The observation that both proteins are found in the nucleus is in agreement with previous genetic data which suggest an interaction between RNA14p and RNA15p. Also the joint nuclear localization is consistent with the biochemical data suggesting a role in polyadenylation. The detection of significant amounts of RNA14p in the cytoplasm opens the possibility of a second function for this protein, either in cytoplasmic regulation of mRNA deadenylation or, more interestingly, in mRNA stability.
RNA14和RNA15最初是通过温度敏感突变鉴定出来的,这些突变会导致在限制温度下聚腺苷酸尾长度和总体mRNA水平迅速下降。我们制备了针对RNA14和RNA15蛋白的抗体,并使用亚细胞分级分离和免疫荧光技术在酵母细胞内定位这些蛋白。RNA14p是一种73 kDa的蛋白,在细胞核和细胞质中均有发现,而RNA15p是一种42 kDa的蛋白,仅在细胞核中检测到。两种蛋白都存在于细胞核中的观察结果与先前的遗传数据一致,这些数据表明RNA14p和RNA15p之间存在相互作用。同样,联合核定位与生化数据一致,表明其在聚腺苷酸化中起作用。在细胞质中检测到大量的RNA14p,这为该蛋白的第二种功能提供了可能性,要么在mRNA去腺苷酸化的细胞质调节中,要么更有趣的是,在mRNA稳定性方面。