A 5′ stem-loop and ribosome binding but not translation are important for the stability of Bacillus subtilis aprE leader mRNA
A 5′ stem-loop and ribosome binding but not translation are important for the stability of Bacillus subtilis aprE leader mRNA
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DOI：
10.1099/00221287-148-6-1795
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发表时间：
2002-06-01
期刊：
影响因子：
2.8
通讯作者：
Rutberg, B
中科院分区：
文献类型：
--
作者：
Hambraeus, G;Karhumaa, K;Rutberg, B
枯草芽孢杆菌aprE前导序列是极端mRNA稳定性的决定因素。作者研究了aprE前导序列的哪些特性赋予mRNA稳定性。在体外和体内分析了aprE前导mRNA的二级结构，并将突变引入aprE前导-lacZ融合体的不同结构域。测定相应转录物的半衰期，并测量β-半乳糖苷酶活性。在5'末端去除茎环结构或降低RBS的强度将半衰期从超过25分钟减少到约5分钟。通过废除起始密码子或创建早期终止密码子来干扰翻译对mRNA稳定性没有或几乎没有影响。作者得出结论，5'茎环和核糖体的结合是aprE前导mRNA稳定所必需的。目前的结果，连同其他一些数据，表明翻译的一个B。枯草杆菌mRNA的稳定性一般不重要;在大肠杆菌中情况似乎不同。进一步得出结论：a B.枯草杆菌RBS不能用于预测相应转录物的稳定性。
The Bacillus subtilis aprE leader is a determinant of extreme MRNA stability. The authors examined what properties of the aprE leader confer stability on an mRNA. The secondary structure of the aprE leader mRNA was analysed in vitro and in vivo, and mutations were introduced into different domains of an aprE leader-lacZ fusion. The half-lives of the corresponding transcripts were determined and,beta-galactosidase activities were measured. Removal of a stem-loop structure at the 5' end or diminishing the strength of the RBS reduced the half-lives from more than 25 min to about 5 min. Interfering with translation by abolishing the start codon or creating an early stop codon had no or little effect on mRNA stability. The authors conclude that a 5' stem-loop and binding of ribosomes are necessary for the stability of aprE leader mRNA. The present results, together with a number of other data, suggest that translation of a B. subtilis mRNA is generally not important for its stability; the situation seems different in Escherichia coli. it is further concluded that the calculated strength of a B. subtilis RBS cannot be used to predict the stability of the corresponding transcript.