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Biochem Biophys Res Commun ; 716: 150009, 2024 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-38697010

RESUMO

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.


Assuntos
DNA Bacteriano , Escherichia coli , Microscopia de Fluorescência , Imagem Individual de Molécula , Microscopia de Fluorescência/métodos , Escherichia coli/genética , Escherichia coli/metabolismo , Imagem Individual de Molécula/métodos , DNA Bacteriano/metabolismo , DNA Bacteriano/genética , Resposta SOS em Genética , DNA de Cadeia Simples/metabolismo , DNA de Cadeia Simples/genética , Proteínas de Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Ligação Proteica , Recombinases Rec A/metabolismo , Recombinases Rec A/genética , Pinças Ópticas
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