Molecular Biology Division, Bhabha Atomic Research Centre, Mumbai, 400085, India.
Life Sciences, Homi Bhabha National Institute, Mumbai, 400094, India.
Mol Microbiol. 2019 Sep;112(3):854-865. doi: 10.1111/mmi.14321. Epub 2019 Jun 19.
The GC-rich genome of Deinococcus radiodurans contains a very high density of putative guanine quadruplex (G4) DNA motifs and its RecQ (drRecQ) was earlier characterized as a 3'→5' dsDNA helicase. We saw that N-Methyl mesoporphyrin IX (NMM), a G4 DNA binding drug affected normal growth as well as the gamma radiation resistance of the wild-type bacterium. Interestingly, NMM treatment and recQ deletion showed additive effect on normal growth but there was no effect of NMM on gamma radiation resistance of recQ mutant. The recombinant drRecQ showed ~400 times higher affinity to G4 DNA (K = 11.74 ± 1.77 nM) as compared to dsDNA (K = 4.88 ± 1.30 µM). drRecQ showed ATP independent helicase function on G4 DNA, which was higher than ATP-dependent helicase activity on dsDNA. Unlike wild-type cells that sparingly stained for G4 structure with Thioflavin T (ThT), recQ mutant showed very high-density of ThT fluorescence foci on DNA indicating an important role of drRecQ in regulation of G4 DNA structure dynamics in vivo. These results together suggested that drRecQ is an ATP independent G4 DNA helicase that plays an important role in the regulation of G4 DNA structure dynamics and its impact on radioresistance in D. radiodurans.
耐辐射球菌的 GC 丰富基因组包含非常高密度的推定鸟嘌呤四链体 (G4) DNA 基序,其 RecQ (drRecQ) 早些时候被表征为 3'→5' dsDNA 解旋酶。我们发现 N-甲基 mesoporphyrin IX (NMM),一种 G4 DNA 结合药物,不仅影响野生型细菌的正常生长,还影响其γ辐射抗性。有趣的是,NMM 处理和 recQ 缺失对正常生长表现出相加效应,但 NMM 对 recQ 突变体的γ辐射抗性没有影响。重组 drRecQ 对 G4 DNA 的亲和力 (~400 倍) 高于 dsDNA (K = 11.74 ± 1.77 nM 对 K = 4.88 ± 1.30 µM)。drRecQ 在 G4 DNA 上表现出 ATP 非依赖性解旋酶功能,高于在 dsDNA 上的 ATP 依赖性解旋酶活性。与用噻唑橙 (ThT) 很少染色 G4 结构的野生型细胞不同,recQ 突变体在 DNA 上显示出非常高密度的 ThT 荧光焦点,表明 drRecQ 在体内 G4 DNA 结构动力学的调节中起着重要作用。这些结果共同表明,drRecQ 是一种 ATP 非依赖性 G4 DNA 解旋酶,在 G4 DNA 结构动力学的调节及其对耐辐射球菌的辐射抗性的影响中起着重要作用。