Wang S S, Stanford D R, Silvers C D, Hopper A K
Department of Biological Chemistry, Milton S. Hershey Medical Center, Pennsylvania State University, Hershey 17033.
Mol Cell Biol. 1992 Jun;12(6):2633-43. doi: 10.1128/mcb.12.6.2633-2643.1992.
STP1 is an unessential yeast gene involved in the removal of intervening sequences from some, but not all, families of intervening sequence-containing pre-tRNAs. Previously, we proposed that STP1 might encode a product that generates pre-tRNA conformations efficiently recognized by tRNA-splicing endonuclease. To test the predictions of this model, we have undertaken a molecular analysis of the STP1 gene and its products. The STP1 locus is located on chromosome IV close to at least two other genes involved in RNA splicing: PRP3 and SPP41. The STP1 open reading frame (ORF) could encode a peptide of 64,827 Da; however, inspection of putative transcriptional and translational regulatory signals and mapping of the 5' ends of mRNA provide evidence that translation of the STP1 ORF usually initiates at a second AUG to generate a protein of 58,081 Da. The STP1 ORF contains three putative zinc fingers. The first of these closely resembles both the DNA transcription factor consensus and the Xenopus laevis p43 RNA-binding protein consensus. The third motif more closely resembles the fingers found in spliceosomal proteins. Employing antisera to the endogenous STP1 protein and to STP1-LacZ fusion proteins, we show that the STP1 protein is localized to nuclei. The presence of zinc finger motifs and the nuclear location of the STP1 protein support the model that this gene product is involved directly in pre-tRNA splicing.
STP1是酵母中的一个非必需基因,参与从部分(而非全部)含居间序列的前体tRNA家族中去除居间序列。此前,我们提出STP1可能编码一种产物,该产物能高效生成可被tRNA剪接内切酶识别的前体tRNA构象。为了验证该模型的预测,我们对STP1基因及其产物进行了分子分析。STP1基因座位于第四条染色体上,靠近至少另外两个参与RNA剪接的基因:PRP3和SPP41。STP1开放阅读框(ORF)可编码一个64,827道尔顿的肽段;然而,对推定的转录和翻译调控信号的检查以及mRNA 5'端的定位提供了证据,表明STP1 ORF的翻译通常在第二个AUG起始,生成一个58,081道尔顿的蛋白质。STP1 ORF包含三个推定的锌指结构。其中第一个与DNA转录因子共有序列以及非洲爪蟾p43 RNA结合蛋白共有序列都非常相似。第三个基序更类似于剪接体蛋白中的锌指结构。利用针对内源性STP1蛋白和STP1-LacZ融合蛋白的抗血清,我们发现STP1蛋白定位于细胞核。锌指基序的存在以及STP1蛋白的核定位支持了该基因产物直接参与前体tRNA剪接的模型。