Enwerem Isioma I, Wu Guowei, Yu Yi Tao, Hebert Michael D
Department of Biochemistry, The University of Mississippi Medical Center, Jackson, Mississippi 39216-4505, United States of America.
Department of Biochemistry and Biophysics, The University of Rochester Medical Center, Rochester, New York 14642, United States of America.
PLoS One. 2015 Apr 13;10(4):e0122348. doi: 10.1371/journal.pone.0122348. eCollection 2015.
Small nuclear ribonucleoproteins (snRNPs), which are required for pre-mRNA splicing, contain extensively modified snRNA. Small Cajal body-specific ribonucleoproteins (scaRNPs) mediate these modifications. It is unknown how the box C/D class of scaRNPs localizes to Cajal Bodies (CBs). The processing of box C/D scaRNA is also unclear. Here, we explore the processing of box C/D scaRNA 2 and 9 by coilin. We also broaden our investigation to include WRAP53 and SMN, which accumulate in CBs, play a role in RNP biogenesis and associate with coilin. These studies demonstrate that the processing of an ectopically expressed scaRNA2 is altered upon the reduction of coilin, WRAP53 or SMN, but the extent and direction of this change varies depending on the protein reduced. We also show that box C/D scaRNP activity is reduced in a cell line derived from coilin knockout mice. Collectively, the findings presented here further implicate coilin as being a direct participant in the formation of box C/D scaRNPs, and demonstrate that WRAP53 and SMN may also play a role, but the activity of these proteins is divergent to coilin.
小核核糖核蛋白(snRNPs)是前体mRNA剪接所必需的,包含经过广泛修饰的snRNA。小 Cajal 体特异性核糖核蛋白(scaRNPs)介导这些修饰。目前尚不清楚 C/D 盒类 scaRNPs 如何定位于 Cajal 体(CBs)。C/D 盒 scaRNA 的加工过程也不清楚。在这里,我们探索了卷曲螺旋蛋白(coilin)对 C/D 盒 scaRNA 2 和 9 的加工。我们还将研究范围扩大到包括 WRAP53 和运动神经元存活蛋白(SMN),它们在 Cajal 体中积累,在核糖核蛋白生物合成中发挥作用,并与卷曲螺旋蛋白相关联。这些研究表明,在卷曲螺旋蛋白、WRAP53 或 SMN 减少时,异位表达的 scaRNA2 的加工会发生改变,但这种变化的程度和方向因减少的蛋白质而异。我们还表明,在源自卷曲螺旋蛋白基因敲除小鼠的细胞系中,C/D 盒 scaRNP 活性降低。总的来说,这里呈现的研究结果进一步表明卷曲螺旋蛋白是 C/D 盒 scaRNPs 形成的直接参与者,并证明 WRAP53 和 SMN 也可能发挥作用,但这些蛋白质的活性与卷曲螺旋蛋白不同。