Xue D, Rubinson D A, Pannone B K, Yoo C J, Wolin S L
Departments of Cell Biology and Molecular Biophysics and Biochemistry, Howard Hughes Medical Institute, Yale University School of Medicine, 295 Congress Avenue, New Haven, CT 06536, USA.
EMBO J. 2000 Apr 3;19(7):1650-60. doi: 10.1093/emboj/19.7.1650.
In all eukaryotic nuclei, the La autoantigen binds nascent RNA polymerase III transcripts, stabilizing these RNAs against exonucleases. Here we report that the La protein also functions in the assembly of certain RNA polymerase II-transcribed RNAs into RNPs. A mutation in a core protein of the spliceosomal snRNPs, Smd1p, causes yeast cells to require the La protein Lhp1p for growth at low temperatures. Precursors to U1, U2, U4 and U5 RNAs are bound by Lhp1p in both wild-type and mutant cells. At the permissive temperature, smd1-1 cells contain higher levels of stable U1 and U5 snRNPs when Lhp1p is present. At low temperatures, Lhp1p becomes essential for the accumulation of U4/U6 snRNPs and for cell viability. When U4 RNA is added to extracts, the pre-U4 RNA, but not the mature RNA, is bound by Smd1p. These results suggest that, by stabilizing a 3'-extended form of U4 RNA, Lhp1p facilitates efficient Sm protein binding, thus assisting formation of the U4/U6 snRNP.
在所有真核细胞核中,La自身抗原会结合新生的RNA聚合酶III转录本,使这些RNA免受核酸外切酶的降解而保持稳定。在此我们报告,La蛋白在某些RNA聚合酶II转录的RNA组装成核糖核蛋白颗粒(RNP)的过程中也发挥作用。剪接体小核核糖核蛋白(snRNP)的核心蛋白Smd1p发生突变,会导致酵母细胞在低温下生长需要La蛋白Lhp1p。在野生型和突变型细胞中,U1、U2、U4和U5 RNA的前体均会与Lhp1p结合。在允许温度下,当存在Lhp1p时,smd1-1细胞含有更高水平的稳定U1和U5 snRNP。在低温下,Lhp1p对于U4/U6 snRNP的积累和细胞活力变得至关重要。当将U4 RNA添加到提取物中时,前体U4 RNA而非成熟RNA会与Smd1p结合。这些结果表明,通过稳定U4 RNA的3'延伸形式,Lhp1p促进了Sm蛋白的有效结合,从而协助U4/U6 snRNP的形成。