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来自裂殖酵母粟酒裂殖酵母的DNA连接酶基因CDC17的分子特征分析。

Molecular characterisation of the DNA ligase gene, CDC17, from the fission yeast Schizosaccharomyces pombe.

作者信息

Barker D G, White J H, Johnston L H

出版信息

Eur J Biochem. 1987 Feb 2;162(3):659-67. doi: 10.1111/j.1432-1033.1987.tb10688.x.

Abstract

We have sequenced a 4200-base-pair fragment of Schizosaccharomyces pombe DNA which encompasses the entire DNA ligase gene, CDC17. S1 mapping has enabled us to identify two small introns (40 and 62 nucleotides) at the 5' end of the coding region of the gene and their 3' internal conserved sequences match the CTRAY consensus found in other S. pombe introns. The major transcription initiation and 3' polyadenylation sites have been mapped and are preceded by higher eukaryotic-like TATA and AATAAA sequences respectively. Furthermore, the CDC17 mRNA carries a poly(A) tail whose length (approximately 250 nucleotides) is typical of that found in higher eukaryotic mRNAs, and is in contrast to the much shorter polyadenylated sequences found for the mRNAs of the budding yeast, Saccharomyces cerevisiae. The deduced amino acid sequence of the S. pombe DNA ligase predicts a protein of 86182 daltons, and an overall 53% homology with the same enzyme from S. cerevisiae. In particular, a stretch of 24 amino acids with 100% sequence homology spans the putative ATP-binding region which is also conserved in T4 and T7 bacteriophage DNA ligases.

摘要

我们对粟酒裂殖酵母DNA的一个4200个碱基对的片段进行了测序,该片段包含整个DNA连接酶基因CDC17。S1图谱分析使我们能够在该基因编码区的5'端识别出两个小内含子(40和62个核苷酸),并且它们的3'内部保守序列与在其他粟酒裂殖酵母内含子中发现的CTRAY共有序列相匹配。主要转录起始位点和3'聚腺苷酸化位点已被定位,并且分别在类似高等真核生物的TATA和AATAAA序列之前。此外,CDC17 mRNA带有一个聚(A)尾,其长度(约250个核苷酸)是高等真核生物mRNA中典型的长度,这与在芽殖酵母酿酒酵母的mRNA中发现的短得多的聚腺苷酸化序列形成对比。粟酒裂殖酵母DNA连接酶的推导氨基酸序列预测出一个86182道尔顿的蛋白质,并且与来自酿酒酵母的相同酶具有53%的总体同源性。特别是,一段24个氨基酸的序列具有100%的序列同源性,跨越了推定的ATP结合区域,该区域在T4和T7噬菌体DNA连接酶中也保守。

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