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 article

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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constrictions
microscopy
proteins
fluorescence
bacteriophages
deoxyribonucleic acid
progeny
plasmids
cells
bacteria
enzymes
breakdown
degradation

Bibliographical note

Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI.

Cite this

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title = "Fluorescence microscopy study of the effect of Esp1396I restriction-modification system proteins concentrations on protection against lambda phage.",
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.",
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Fluorescence microscopy study of the effect of Esp1396I restriction-modification system proteins concentrations on protection against lambda phage. / Smirnov, S. V.; Morozova, N. E.; Khodorkovskii, M. A.; Severinov, K. V.

In: Journal of Physics: Conference Series, Vol. 1135, No. 1, 012016, 20.12.2018.

Research output: Contribution to journalConference article

TY - JOUR

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AU - Smirnov, S. V.

AU - Morozova, N. E.

AU - Khodorkovskii, M. A.

AU - Severinov, K. V.

N1 - Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI.

PY - 2018/12/20

Y1 - 2018/12/20

N2 - 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.

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