Potnis Akhilesh A, Raghavan Prashanth S, Shelke Ashwini, Nikam T D, Rajaram Hema
Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai-400085, India.
Department of Botany, Savitribai Phule Pune University, Ganeshkhind, Pune-411007, India.
FEMS Microbiol Lett. 2017 Jan;364(1). doi: 10.1093/femsle/fnw279. Epub 2016 Dec 8.
Anabaena PCC7120 has two annotated toxin-antitoxin systems: MazEF and HicAB. Overexpression of either of the toxins severely inhibited the growth of Escherichia coli BL21(plysS)(DE3). Of the two Anabaena toxins, MazF exhibited higher toxicity than HicA as evidenced by (i) 100-fold lower viability upon overexpression of MazF compared to HicA; (ii) complete loss of cell viability within 1 h of induction of MazF expression, as against >10 colony forming units mL in case of HicA; (iii) inability to maintain the MazF overexpressing plasmid in E. coli cells; and (iv) neutralisation of the toxin was effective at the molar ratio of 1:1.9 for MazF:MazE and 13:1 for HicA:HicB, indicating higher antitoxin requirement for neutralisation of MazF. The growth inhibitory effect of MazF was found to be higher in lag phase cultures compared to mid-logarithmic phase cultures of E. coli, while the reverse was true for HicA. The results suggest possible distinct roles for MazEF and HicAB systems of Anabaena.
鱼腥藻PCC7120有两个已注释的毒素-抗毒素系统:MazEF和HicAB。任一毒素的过表达都会严重抑制大肠杆菌BL21(plysS)(DE3)的生长。在这两种鱼腥藻毒素中,MazF表现出比HicA更高的毒性,这体现在以下方面:(i)与HicA相比,MazF过表达时的存活率低100倍;(ii)诱导MazF表达后1小时内细胞活力完全丧失,而HicA的情况是每毫升有>10个菌落形成单位;(iii)无法在大肠杆菌细胞中维持MazF过表达质粒;(iv)毒素中和时,MazF与MazE的摩尔比为1:1.9,HicA与HicB的摩尔比为13:1时有效,这表明中和MazF需要更高的抗毒素量。结果发现,与大肠杆菌对数中期培养物相比,MazF对滞后期培养物的生长抑制作用更高,而HicA的情况则相反。这些结果表明鱼腥藻的MazEF和HicAB系统可能具有不同的作用。