Vázquez-Cruz C, Olmedo-Alvarez G
Departamento de Ingeniería Genética, CINVESTAV IPN, Unidad Irapuato, Guanajuato, México.
J Bacteriol. 1997 Oct;179(20):6341-8. doi: 10.1128/jb.179.20.6341-6348.1997.
We undertook the study of the decay process of the cry1Aa mRNA of Bacillus thuringiensis expressed in B. subtilis. The cry1Aa transcript is a 3.7-kb mRNA expressed during sporulation whose transcriptional control has previously been studied in both B. subtilis and B. thuringiensis. We found that the cry1Aa mRNA has a half-life of around 9 min and that its decay occurs through endoribonucleolytic cleavages which result in three groups of high-molecular-weight mRNA intermediates ranging in size from 2.7 to 0.5 kb. A comparative study carried out with Escherichia coli showed a similar pattern of degradation intermediates. Primer extension analysis carried out on RNA from B. subtilis revealed that most cleavages occur within two regions located toward the 5' and 3' ends of the mRNA. The most prominent processing site observed for the cry1Aa mRNA isolated from B. subtilis is only two bases away from that occurring on RNA isolated from E. coli. Most cleavage sites occur at seemingly single-stranded RNA segments rich in A and U nucleotides, suggesting that a common and conserved mechanism may process the cry1Aa mRNA.
我们对在枯草芽孢杆菌中表达的苏云金芽孢杆菌cry1Aa mRNA的衰变过程进行了研究。cry1Aa转录本是一种在芽孢形成过程中表达的3.7 kb mRNA,其转录调控此前已在枯草芽孢杆菌和苏云金芽孢杆菌中进行过研究。我们发现cry1Aa mRNA的半衰期约为9分钟,其衰变通过核糖核酸内切酶切割发生,这导致产生三组高分子量mRNA中间体,大小从2.7 kb到0.5 kb不等。与大肠杆菌进行的比较研究显示出类似的降解中间体模式。对来自枯草芽孢杆菌的RNA进行的引物延伸分析表明,大多数切割发生在mRNA 5'端和3'端附近的两个区域内。从枯草芽孢杆菌分离的cry1Aa mRNA观察到的最突出加工位点与从大肠杆菌分离的RNA上的位点仅相差两个碱基。大多数切割位点出现在富含A和U核苷酸的看似单链RNA片段上,这表明可能存在一种共同且保守的机制来加工cry1Aa mRNA。