Antimicrobial resistance threatens the eradication of infectious diseases and impairs the efficacy of available therapeutics. The bacterial SOS pathway is a conserved response triggered by genotoxic stresses and represents one of the principal mechanisms that lead to resistance. The RecA recombinase acts as a DNA-damage sensor inducing the autoproteolysis of the transcriptional repressor LexA, thereby derepressing SOS genes that mediate DNA repair, survival to chemotherapy, and hypermutation. The inhibition of such pathway represents a promising strategy for delaying the evolution of antimicrobial resistance. We report the identification, via llama immunization and phage display, of nanobodies that bind LexA with sub-micromolar affinity and block autoproteolysis, repressing SOS response in Escherichia coli. Biophysical characterization of nanobody-LexA complexes revealed that they act by trapping LexA in an inactive conformation and interfering with RecA engagement. Our studies pave the way to the development of new-generation antibiotic adjuvants for the treatment of bacterial infections.

Nanobodies targeting LexA autocleavage disclose a novel suppression strategy of SOS-response pathway / Maso, Lorenzo; Vascon, Filippo; Chinellato, Monica; Goormaghtigh, Frédéric; Bellio, Pierangelo; Campagnaro, Enrica; Van Melderen, Laurence; Ruzzene, Maria; Pardon, Els; Angelini, Alessandro; Celenza, Giuseppe; Steyaert, Jan; Tondi, Donatella; Cendron, Laura. - In: STRUCTURE. - ISSN 0969-2126. - (2022), pp. 1-40. [10.1016/j.str.2022.09.004]

Nanobodies targeting LexA autocleavage disclose a novel suppression strategy of SOS-response pathway

Tondi, Donatella;
2022-01-01

Abstract

Antimicrobial resistance threatens the eradication of infectious diseases and impairs the efficacy of available therapeutics. The bacterial SOS pathway is a conserved response triggered by genotoxic stresses and represents one of the principal mechanisms that lead to resistance. The RecA recombinase acts as a DNA-damage sensor inducing the autoproteolysis of the transcriptional repressor LexA, thereby derepressing SOS genes that mediate DNA repair, survival to chemotherapy, and hypermutation. The inhibition of such pathway represents a promising strategy for delaying the evolution of antimicrobial resistance. We report the identification, via llama immunization and phage display, of nanobodies that bind LexA with sub-micromolar affinity and block autoproteolysis, repressing SOS response in Escherichia coli. Biophysical characterization of nanobody-LexA complexes revealed that they act by trapping LexA in an inactive conformation and interfering with RecA engagement. Our studies pave the way to the development of new-generation antibiotic adjuvants for the treatment of bacterial infections.
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Nanobodies targeting LexA autocleavage disclose a novel suppression strategy of SOS-response pathway / Maso, Lorenzo; Vascon, Filippo; Chinellato, Monica; Goormaghtigh, Frédéric; Bellio, Pierangelo; Campagnaro, Enrica; Van Melderen, Laurence; Ruzzene, Maria; Pardon, Els; Angelini, Alessandro; Celenza, Giuseppe; Steyaert, Jan; Tondi, Donatella; Cendron, Laura. - In: STRUCTURE. - ISSN 0969-2126. - (2022), pp. 1-40. [10.1016/j.str.2022.09.004]
Maso, Lorenzo; Vascon, Filippo; Chinellato, Monica; Goormaghtigh, Frédéric; Bellio, Pierangelo; Campagnaro, Enrica; Van Melderen, Laurence; Ruzzene, Maria; Pardon, Els; Angelini, Alessandro; Celenza, Giuseppe; Steyaert, Jan; Tondi, Donatella; Cendron, Laura
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11380/1289685
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