Hue K K, Bechhofer D H
Department of Biochemistry, Mount Sinai School of Medicine, New York, New York 10029-6574.
J Bacteriol. 1991 Jun;173(12):3732-40. doi: 10.1128/jb.173.12.3732-3740.1991.
Induction of translation of the ermC gene product in Bacillus subtilis occurs upon exposure to erythromycin and is a result of ribosome stalling in the ermC leader peptide coding sequence. Another result of ribosome stalling is stabilization of ermC mRNA. The effect of leader RNA secondary structure, methylase translation, and leader peptide translation on induced ermC mRNA stability was examined by constructing various mutations in the ermC leader region. Analysis of deletion mutations showed that ribosome stalling causes induction of ermC mRNA stability in the absence of methylase translation and ermC leader RNA secondary structure. Furthermore, deletions that removed much of the leader peptide coding sequence had no effect on induced ermC mRNA stability. A leader region mutation was constructed such that ribosome stalling occurred in a position upstream of the natural stall site, resulting in induced mRNA stability without induction of translation. This mutation was used to measure the effect of mRNA stabilization on ermC gene expression.
在枯草芽孢杆菌中,当暴露于红霉素时,ermC基因产物的翻译被诱导,这是核糖体在ermC前导肽编码序列中停滞的结果。核糖体停滞的另一个结果是ermC mRNA的稳定。通过在ermC前导区域构建各种突变,研究了前导RNA二级结构、甲基化酶翻译和前导肽翻译对诱导的ermC mRNA稳定性的影响。缺失突变分析表明,在没有甲基化酶翻译和ermC前导RNA二级结构的情况下,核糖体停滞会导致ermC mRNA稳定性的诱导。此外,去除大部分前导肽编码序列的缺失对诱导的ermC mRNA稳定性没有影响。构建了一个前导区域突变,使得核糖体停滞发生在天然停滞位点上游的位置,导致mRNA稳定性的诱导而不诱导翻译。该突变用于测量mRNA稳定对ermC基因表达的影响。