Department of Microbiology, University of Gdansk, 80-822 Gdansk, Kladki 24, Poland.
J Microbiol Biotechnol. 2010 Jan;20(1):30-8.
The contribution of a type II restriction-modification system (R-M system) to genome integrity and cell viability was investigated. We established experimental conditions which enabled the achievement of hemimethylated and unmethylated states for the specific bases of the recognition sequences of the host's DNA. To achieve this, we constructed the MboII R-M system containing only one (i.e. M2.MboII) out of two functional MboII methyltransferases found in Moraxella bovis. Using the incomplete R-M system we were able to perturb the balance between methylation and restriction in an inducible manner. We demonstrate that upon the SOS-induced DNA repair in the mitomycin C treated cells, restriction significantly reduces cell viability. Similar results for the well-studied wild type EcoRI R-M system, expressed constitutively in Escherichia coli, were obtained. Our data provide further insights into the benefits and disadvantages of maintaining of a type II R-M system, highlighting its impact on host cell fitness.
研究了 II 型限制-修饰(R-M)系统对基因组完整性和细胞活力的贡献。我们建立了实验条件,使宿主 DNA 的识别序列的特定碱基能够达到半甲基化和非甲基化状态。为此,我们构建了仅包含莫拉氏菌中发现的两个功能性 MboII 甲基转移酶之一(即 M2.MboII)的 MboII R-M 系统。使用不完整的 R-M 系统,我们能够以诱导的方式改变甲基化和限制之间的平衡。我们证明,在丝裂霉素 C 处理的细胞中诱导 SOS 修复时,限制会显著降低细胞活力。在组成型表达于大肠杆菌中的研究更为充分的野生型 EcoRI R-M 系统中也获得了类似的结果。我们的数据进一步深入了解了维持 II 型 R-M 系统的利弊,强调了其对宿主细胞适应性的影响。