Dupuis P
Centre de Génétique Moléculaire, Université Pierre et Marie Curie, Gifsur-Yvette, France.
EMBO J. 1992 Oct;11(10):3713-9. doi: 10.1002/j.1460-2075.1992.tb05456.x.
In the ciliate Paramecium tetraurelia, the analysis of the tubulin gene family has revealed the existence of four alpha and three beta genes. We show here that the coding sequence of the first beta-tubulin gene to be cloned and sequenced is interrupted by two short non-coding sequences of 27 bp each, which present at their extremities the pairs GT/AG, characteristic of eukaryotic pre-mRNA introns, and the internal pentanucleotide TTAAT, consensual in Tetrahymena introns. We demonstrate by PCR experiments that the three macronuclear beta-tubulin genes contain these sequences in similar positions, thereby ruling out the possibility that these sequences are ciliate IES present in the micronucleus and eliminated in the transcriptionally active macronucleus. S1 mapping analysis and mRNA sequencing show that the sequences are absent from the beta-tubulin transcripts. These sequences are the first introns described in protein encoding genes in P. tetraurelia and the shortest known introns altogether.
在纤毛虫四膜虫中,对微管蛋白基因家族的分析揭示了四个α基因和三个β基因的存在。我们在此表明,首个被克隆和测序的β微管蛋白基因的编码序列被两个各为27 bp的短非编码序列打断,这两个序列在其末端呈现出GT/AG对,这是真核生物前体mRNA内含子的特征,并且还含有内部五核苷酸TTAAT,这在嗜热四膜虫内含子中是一致的。我们通过PCR实验证明,三个大核β微管蛋白基因在相似位置含有这些序列,从而排除了这些序列是存在于小核中并在转录活跃的大核中被消除的纤毛虫内部消除序列(IES)的可能性。S1图谱分析和mRNA测序表明,β微管蛋白转录本中不存在这些序列。这些序列是在四膜虫蛋白质编码基因中描述的首个内含子,也是目前已知最短的内含子。