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Biochem Biophys Res Commun ; 716: 150009, 2024 Jul 05.
Artículo en Inglés | MEDLINE | ID: mdl-38697010

RESUMEN

The SOS response is a condition that occurs in bacterial cells after DNA damage. In this state, the bacterium is able to reсover the integrity of its genome. Due to the increased level of mutagenesis in cells during the repair of DNA double-strand breaks, the SOS response is also an important mechanism for bacterial adaptation to the antibiotics. One of the key proteins of the SOS response is the SMC-like protein RecN, which helps the RecA recombinase to find a homologous DNA template for repair. In this work, the localization of the recombinant RecN protein in living Escherichia coli cells was revealed using fluorescence microscopy. It has been shown that the RecN, outside the SOS response, is predominantly localized at the poles of the cell, and in dividing cells, also localized at the center. Using in vitro methods including fluorescence microscopy and optical tweezers, we show that RecN predominantly binds single-stranded DNA in an ATP-dependent manner. RecN has both intrinsic and single-stranded DNA-stimulated ATPase activity. The results of this work may be useful for better understanding of the SOS response mechanism and homologous recombination process.


Asunto(s)
ADN Bacteriano , Escherichia coli , Microscopía Fluorescente , Imagen Individual de Molécula , Microscopía Fluorescente/métodos , Escherichia coli/genética , Escherichia coli/metabolismo , Imagen Individual de Molécula/métodos , ADN Bacteriano/metabolismo , ADN Bacteriano/genética , Respuesta SOS en Genética , ADN de Cadena Simple/metabolismo , ADN de Cadena Simple/genética , Proteínas de Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Unión Proteica , Rec A Recombinasas/metabolismo , Rec A Recombinasas/genética , Pinzas Ópticas
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