Danilova, Y.A.; Belousova, V.V.; Moiseenko, A.V.; Vishnyakov, I.E.; Yakunina, M.V.; Sokolova, O.S. Maturation of Pseudo-Nucleus Compartment in P. aeruginosa, Infected with Giant phiKZ Phage. Viruses2020, 12, 1197.
Danilova, Y.A.; Belousova, V.V.; Moiseenko, A.V.; Vishnyakov, I.E.; Yakunina, M.V.; Sokolova, O.S. Maturation of Pseudo-Nucleus Compartment in P. aeruginosa, Infected with Giant phiKZ Phage. Viruses 2020, 12, 1197.
Bacteria develop various defense mechanisms against stresses, including the bacteriophage infection. The giant phiKZ phage infection induced the appearance of a pseudo-nucleus inside the bacterial cytoplasm. Here, we used FISH, electron tomography and analytical electron microscopy to study the morphology of this unique nucleus-like shell and to demonstrate the distribution of phiKZ and bacterial DNA in infected P. aeruginosa cells. The maturation of the pseudo-nucleus was traced in short intervals for 40 min after infection. This study was accompanied by the identification of phiKZ and bacterial DNA by real-time RCR. We demonstrated that phage DNA that isolated from the cytoplasm during all infection stages were compacted within the pseudo-nucleus in a specific structure. Bacterial DNA was diminished in the course of infection, but did not completely degrade until at least 40 min after phage application. The content of the total phage DNA, on the other hand, increased. EDX analysis confirmed these results and revealed that, during the infection, Sulfur content in the bacterial cytoplasm increased, which suggests the increase of DNA-binding Met-reach proteins synthesis, which could protect bacterial DNA from stress.
giant phage; phiKZ; Pseudomonas aeruginosa; nucleoid; pseudo-nucleus; analytical electron microscopy; electron tomography; fluorescent in situ hybridization; stress response
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