Fluorescence microscopy study of the effect of Esp1396I restriction-modification system proteins concentrations on protection against lambda phage.

S. V. Smirnov, N. E. Morozova, M. A. Khodorkovskii, K. V. Severinov

    Research output: Contribution to journalConference articlepeer-review

    Abstract

    Restriction-modification systems are among wide-spread mechanisms protecting bacteria from foreign DNA, such as DNA of bacteriophages and plasmids. The action of such systems is based on the presence of two enzymes - methyltransferase and restriction endonuclease, first one protects host DNA from degradation by modification and second one cleaves foreign unmodified DNA. In some cases bacteriophage DNA is modified faster than being degraded by restriction endonuclease. This process, called breakdown, leads to formation of modified bacteriophage progeny which is not sensitive to the action of restriction endonuclease and can eliminate whole "protected" bacterial population. There is a hypothesis which assumes that the overcoming of the protective action of restriction-modification system by a bacteriophage depends on the relative concentrations of the restriction-modification system proteins. To check this assumption we decided to perform fluorescence microscopy of individual living E.coli cells. For this purposes we use fluorescently-labelled type II Esp1396I restriction-modification system and also create an artificial system which allows to regulate the ratio of restriction-modification system proteins through the additional restriction endonuclease expression. Preliminary results showed an increase in protection with an increase in the restriction endonuclease concentration in cells.

    Original languageEnglish
    Article number012016
    JournalJournal of Physics: Conference Series
    Volume1135
    Issue number1
    DOIs
    Publication statusPublished - 20 Dec 2018
    EventInternational Conference PhysicA.SPb 2018 - Saint Petersburg, Russian Federation
    Duration: 23 Oct 201825 Oct 2018

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