Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tokyo, 135-0064, Japan.
Department of Applied Biological Science, Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa, 761-0795, Japan.
FEMS Microbiol Ecol. 2022 Apr 21;98(4). doi: 10.1093/femsec/fiac037.
The ability to degrade exogenous compounds is acquired by adaptive processes of microorganisms when they are exposed to compounds that are foreign to their existing enzyme systems. Previously, we reported that simultaneous point mutations and mobile genetic elements cause the evolution and optimization of the degradation systems for aromatic compounds. In the present study, we propose another element with this role-tandem repeats. The novel metagenomic tandem repeat (MTR) sequence T(G/A)ACATG(A/C)T was identified in the 5'-untranslated regions of catechol 2,3-dioxygenase (C23O)-encoding genes by metagenomic analysis. Recombinant Escherichia coli carrying a C23O gene with various numbers of MTRs exhibited increased C23O protein expression and enzyme activity compared with cells expressing the C23O gene without MTRs. Real-time reverse transcription PCR showed that changes in the numbers of MTRs affected the levels of detectable C23O mRNA in the E. coli host. Furthermore, the mRNAs transcribed from C23O genes containing various numbers of MTRs had longer half-lives than those transcribed from a C23O gene without MTRs. Thus, MTRs would affect the translation efficiency of the gene expression system. MTRs may change the expression levels of their downstream genes for adaptation to a fluctuating environment.
当微生物暴露于其现有酶系统以外的化合物时,它们会通过适应性过程获得降解外来化合物的能力。以前,我们曾报道过,点突变和移动遗传元件的同时发生导致了芳香族化合物降解系统的进化和优化。在本研究中,我们提出了另一个具有这种作用的元件——串联重复序列。通过宏基因组分析,在儿茶酚 2,3-双加氧酶(C23O)编码基因的 5'-非翻译区中发现了新型的微生物组串联重复(MTR)序列 T(G/A)ACATG(A/C)T。与没有 MTR 的 C23O 基因表达细胞相比,携带带有不同数量 MTR 的 C23O 基因的重组大肠杆菌表现出更高的 C23O 蛋白表达和酶活性。实时反转录 PCR 显示,MTR 数量的变化影响了大肠杆菌宿主中可检测到的 C23O mRNA 水平。此外,从含有不同数量 MTR 的 C23O 基因转录的 mRNA 比从没有 MTR 的 C23O 基因转录的 mRNA 具有更长的半衰期。因此,MTR 可能会影响基因表达系统的翻译效率。MTR 可能会改变其下游基因的表达水平,以适应波动的环境。