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首页> 外文期刊>Molecular and Cellular Biology >A 15-base-pair element activates the SPS4 gene midway through sporulation in Saccharomyces cerevisiae.
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A 15-base-pair element activates the SPS4 gene midway through sporulation in Saccharomyces cerevisiae.

机译：一个15个碱基对的元件在酿酒酵母中通过孢子形成中途激活了SPS4基因。

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Sporulation of the yeast Saccharomyces cerevisiae represents a simple developmental process in which the events of meiosis and spore wall formation are accompanied by the sequential activation of temporally distinct classes of genes. In this study, we have examined expression of the SPS4 gene, which belongs to a group of genes that is activated midway through sporulation. We mapped the upstream boundary of the regulatory region of SPS4 by monitoring the effect of sequential deletions of 5'-flanking sequence on expression of plasmid-borne versions of SPS4 introduced into a MATa/MAT alpha delta sps4/delta sps4 strain. This analysis indicated that the 5' boundary of the regulatory region was within 50 bp of the putative TATA box of the gene. By testing various oligonucleotides that spanned this boundary and the downstream sequence for their ability to activate expression of a heterologous promoter, we found that a 15-bp sequence sufficed to act as a sporulation-specific upstream activation sequence. This 15-bp fragment, designated UASSPS4, activated expression of a CYC1-lacZ reporter gene midway through sporulation and was equally active in both orientations. Extending the UAS fragment to include the adjacent 14-bp enhanced its activity 10-fold. We show that expression of SPS4 is regulated in a manner distinct from that of early meiotic genes: mutation of UME6 did not lead to vegetative expression of SPS4, and sporulation-specific expression was delayed by mutation of IME2. In vivo and in vitro assays suggested that a factor present in vegetative cells bind to the UASSPS4 element. We speculate that during sporulation this factor is modified to serve as an activator of the SPS4 gene or, alternatively, that it recruits an activator to the promoter.
机译：酵母酿酒酵母的孢子形成代表一个简单的发育过程，其中减数分裂和孢子壁形成的事件伴随着时间上不同类别的基因的顺序激活。在这项研究中，我们检查了SPS4基因的表达，该基因属于在孢子形成过程中被激活的一组基因。我们通过监测5'侧翼序列的连续缺失对引入MATa / MATαdelta sps4 / delta sps4菌株的SPS4质粒携带型表达的影响，对SPS4调控区的上游边界作图。该分析表明调节区的5'边界在该基因的假定的TATA盒的50bp之内。通过测试跨越此边界和下游序列的各种寡核苷酸激活异源启动子表达的能力，我们发现一个15 bp的序列足以充当孢子形成特异性上游激活序列。这个15 bp的片段（称为UASSPS4）在孢子形成中途激活了CYC1-lacZ报告基因的表达，并且在两个方向上均具有相同的活性。扩展UAS片段使其包含相邻的14 bp可以将其活性提高10倍。我们显示，SPS4的表达受到与早期减数分裂基因不同的调控：UME6的突变不会导致SPS4的营养表达，而孢子特异性表达会因IME2的突变而延迟。体内和体外测定表明，营养细胞中存在的因子与UASSPS4元素结合。我们推测在孢子形成过程中，该因子被修饰以充当SPS4基因的激活子，或者，它可以将激活子招募至启动子。

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