Temporal dynamics of methyltransferase and restriction endonuclease accumulation in individual cells after introducing a restriction-modification system

Natalia Morozova, Anton Sabantsev, Ekaterina Bogdanova, Yana Fedorova, Anna Maikova, Alexey Vedyaykin, Andjela Rodic, Marko Djordjevic, Mikhail Khodorkovskii, Konstantin Severinov

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Type II restriction-modification (R-M) systems encode a restriction endonuclease that cleaves DNA at specific sites, and a methyltransferase that modifies same sites protecting them from restriction endonuclease cleavage. Type II R-M systems benefit bacteria by protecting them from bacteriophages. Many type II R-M systems are plasmid-based and thus capable of horizontal transfer. Upon the entry of such plasmids into a naïve host with unmodified genomic recognition sites, methyltransferase should be synthesized first and given sufficient time to methylate recognition sites in the bacterial genome before the toxic restriction endonuclease activity appears. Here, we directly demonstrate a delay in restriction endonuclease synthesis after transformation of Escherichia coli cells with a plasmid carrying the Esp1396I type II R-M system, using single-cell microscopy. We further demonstrate that before the appearance of the Esp1396I restriction endonuclease the intracellular concentration of Esp1396I methyltransferase undergoes a sharp peak, which should allow rapid methylation of host genome recognition sites. A mathematical model that satisfactorily describes the observed dynamics of both Esp1396I enzymes is presented. The results reported here were obtained using a functional Esp1396I type II R-M system encoding both enzymes fused to fluorescent proteins. Similar approaches should be applicable to the studies of other R-M systems at single-cell level.

Original languageEnglish (US)
Pages (from-to)790-800
Number of pages11
JournalNucleic acids research
Volume44
Issue number2
DOIs
StatePublished - Jan 29 2016

All Science Journal Classification (ASJC) codes

  • Genetics

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    Morozova, N., Sabantsev, A., Bogdanova, E., Fedorova, Y., Maikova, A., Vedyaykin, A., Rodic, A., Djordjevic, M., Khodorkovskii, M., & Severinov, K. (2016). Temporal dynamics of methyltransferase and restriction endonuclease accumulation in individual cells after introducing a restriction-modification system. Nucleic acids research, 44(2), 790-800. https://doi.org/10.1093/nar/gkv1490