Mita K, Morimyo M, Ito K, Sugaya K, Ebihara K, Hongo E, Higashi T, Hirayama Y, Nakamura Y
Genome Research Group, National Institute of Radiological Sciences, Inage-ku, Chiba, Japan.
Gene. 1997 Mar 18;187(2):259-66. doi: 10.1016/s0378-1119(96)00764-0.
In the course of the Schizosaccharomyces pombe cDNA project, we succeeded in cloning all the genes encoding translation elongation factors EF-1alpha, EF-1beta, EF-1gamma, EF-2 and EF-3. With the exception of the EF-1gamma gene, the nucleotide (nt) sequence of S. pombe elongation factors has not been previously reported. For EF-1alpha, we found three genes whose amino acid (aa) sequences are quite homologous each other (99.5%), but whose 3' untranslated regions (UTRs) are completely different. Southern blot indicated that those three EF-1alpha genes are located at different loci. Northern analysis indicated that one of three EF-1alpha genes was inducible with UV-irradiation, while the level of expression for another of three EF-1alpha genes was repressed by UV and heat-shock (HS) treatments. The aa sequence predicted from the nt sequence of the S. pombe EF-1beta cDNA clone covered almost all the coding sequence (CDS) of EF-1beta except the first methionine which has 55.4% identity with that of S. cerevisiae. We also identified two copies of S. pombe EF-2 genes. Their aa sequences deduced from nt sequences are identical (100%), but they have different 3' UTRs. The location of these two EF-2 genes in different loci was proved by Southern analysis. The S. pombe EF-3 cDNA clone encoded only a third of the CDS from the C-terminal and its deduced aa sequence has a 76% identity with those of other yeasts and fungi.
在粟酒裂殖酵母cDNA项目过程中,我们成功克隆了所有编码翻译延伸因子EF-1α、EF-1β、EF-1γ、EF-2和EF-3的基因。除EF-1γ基因外,粟酒裂殖酵母延伸因子的核苷酸(nt)序列此前尚未见报道。对于EF-1α,我们发现了三个基因,它们的氨基酸(aa)序列彼此非常同源(99.5%),但其3'非翻译区(UTR)完全不同。Southern印迹表明这三个EF-1α基因位于不同位点。Northern分析表明,三个EF-1α基因中的一个可被紫外线照射诱导,而另外三个EF-1α基因中的一个的表达水平在紫外线和热休克(HS)处理后受到抑制。从粟酒裂殖酵母EF-1β cDNA克隆的nt序列预测的aa序列几乎覆盖了EF-1β的所有编码序列(CDS),除了第一个甲硫氨酸,它与酿酒酵母的甲硫氨酸具有55.4%的同一性。我们还鉴定出两个粟酒裂殖酵母EF-2基因拷贝。从nt序列推导的它们的aa序列是相同的(100%),但它们具有不同的3'UTR。Southern分析证明了这两个EF-2基因位于不同位点。粟酒裂殖酵母EF-3 cDNA克隆仅编码来自C末端的三分之一CDS,其推导的aa序列与其他酵母和真菌的aa序列具有76%的同一性。