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一种短的顺式作用DNA序列的特性分析，该序列在酿酒酵母中传递细胞周期阶段依赖性转录。

Characterization of a short, cis-acting DNA sequence which conveys cell cycle stage-dependent transcription in Saccharomyces cerevisiae.

作者信息

McIntosh E M, Atkinson T, Storms R K, Smith M

机构信息

Department of Biochemistry, University of British Columbia, Vancouver, Canada.

出版信息

Mol Cell Biol. 1991 Jan;11(1):329-37. doi: 10.1128/mcb.11.1.329-337.1991.

DOI:10.1128/mcb.11.1.329-337.1991
PMID:1986229
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC359623/
Abstract

Comparison of the 5'-flanking regions of several cell cycle-regulated DNA replication genes of Saccharomyces cerevisiae has revealed the presence of a common sequence, 5'-ACGCGT-3', which is upstream and proximal to mapped transcription initiation sites. This sequence, which is the cleavage site for the restriction endonuclease MluI, is present twice in the upstream region of the yeast thymidylate synthase gene TMP1. Previous studies have implicated these MluI sites as critical components in the cell cycle-dependent transcription of TMP1. In this study, we examined more closely the importance of the ACGCGT sequences for the transcription of this gene. Using site-directed mutagenesis in combination with deletion analysis and subcloning experiments, we found that (i) while both of the TMP1 MluI sites contribute to the total transcription of this gene, the distal site is predominant and (ii) the 9-bp sequence ACGCGTTAA encompassing the distal MluI site exhibits properties of a cell cycle-stage dependent upstream activation sequence element. The results of this study support the notion that the ACGCGT sequence is an integral component of a transcription system which coordinates the cell cycle-dependent expression of DNA replication genes in S. cerevisiae.

摘要

对酿酒酵母几个细胞周期调控的DNA复制基因的5'侧翼区域进行比较后发现，存在一个共同序列5'-ACGCGT-3'，该序列位于已定位的转录起始位点的上游且与之相邻。这个序列是限制性内切酶MluI的切割位点，在酵母胸苷酸合成酶基因TMP1的上游区域出现了两次。先前的研究表明这些MluI位点是TMP1细胞周期依赖性转录的关键组成部分。在本研究中，我们更深入地研究了ACGCGT序列对该基因转录的重要性。通过将定点诱变与缺失分析和亚克隆实验相结合，我们发现：（i）虽然TMP1的两个MluI位点都对该基因的总转录有贡献，但远端位点起主要作用；（ii）包含远端MluI位点的9碱基序列ACGCGTTAA具有细胞周期阶段依赖性上游激活序列元件的特性。本研究结果支持这样一种观点，即ACGCGT序列是一个转录系统的组成部分，该转录系统协调酿酒酵母中DNA复制基因的细胞周期依赖性表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e90/359623/357435c30b6d/molcellb00136-0347-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e90/359623/45cf3646f436/molcellb00136-0344-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e90/359623/51abe7abd4a5/molcellb00136-0344-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e90/359623/78c75b50a0a1/molcellb00136-0345-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e90/359623/d7137be9a308/molcellb00136-0346-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e90/359623/0641a74c2d83/molcellb00136-0346-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e90/359623/357435c30b6d/molcellb00136-0347-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e90/359623/45cf3646f436/molcellb00136-0344-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e90/359623/51abe7abd4a5/molcellb00136-0344-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e90/359623/78c75b50a0a1/molcellb00136-0345-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e90/359623/d7137be9a308/molcellb00136-0346-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e90/359623/0641a74c2d83/molcellb00136-0346-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e90/359623/357435c30b6d/molcellb00136-0347-a.jpg

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