Staphylococcal DNA Repair Is Required for Infection

ABSTRACT To cause infection, Staphylococcus aureus must withstand damage caused by host immune defenses. However, the mechanisms by which staphylococcal DNA is damaged and repaired during infection are poorly understood. Using a panel of transposon mutants, we identified the rexBA operon as being im...

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Autores principales:	Kam Pou Ha, Rebecca S. Clarke, Gyu-Lee Kim, Jane L. Brittan, Jessica E. Rowley, Despoina A. I. Mavridou, Dane Parker, Thomas B. Clarke, Angela H. Nobbs, Andrew M. Edwards
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	respiratory burst oxidative burst DNA damage Enterococcus SOS system Staphylococcus Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/c366ab45cdbd41dbacadbfc7679c2b74
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spelling	oai:doaj.org-article:c366ab45cdbd41dbacadbfc7679c2b742021-11-15T15:55:43ZStaphylococcal DNA Repair Is Required for Infection10.1128/mBio.02288-202150-7511https://doaj.org/article/c366ab45cdbd41dbacadbfc7679c2b742020-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02288-20https://doaj.org/toc/2150-7511ABSTRACT To cause infection, Staphylococcus aureus must withstand damage caused by host immune defenses. However, the mechanisms by which staphylococcal DNA is damaged and repaired during infection are poorly understood. Using a panel of transposon mutants, we identified the rexBA operon as being important for the survival of Staphylococcus aureus in whole human blood. Mutants lacking rexB were also attenuated for virulence in murine models of both systemic and skin infections. We then demonstrated that RexAB is a member of the AddAB family of helicase/nuclease complexes responsible for initiating the repair of DNA double-strand breaks. Using a fluorescent reporter system, we were able to show that neutrophils cause staphylococcal DNA double-strand breaks through reactive oxygen species (ROS) generated by the respiratory burst, which are repaired by RexAB, leading to the induction of the mutagenic SOS response. We found that RexAB homologues in Enterococcus faecalis and Streptococcus gordonii also promoted the survival of these pathogens in human blood, suggesting that DNA double-strand break repair is required for Gram-positive bacteria to survive in host tissues. Together, these data demonstrate that DNA is a target of host immune cells, leading to double-strand breaks, and that the repair of this damage by an AddAB-family enzyme enables the survival of Gram-positive pathogens during infection. IMPORTANCE To cause infection, bacteria must survive attack by the host immune system. For many bacteria, including the major human pathogen Staphylococcus aureus, the greatest threat is posed by neutrophils. These immune cells ingest the invading organisms and try to kill them with a cocktail of chemicals that includes reactive oxygen species (ROS). The ability of S. aureus to survive this attack is crucial for the progression of infection. However, it was not clear how the ROS damaged S. aureus and how the bacterium repaired this damage. In this work, we show that ROS cause breaks in the staphylococcal DNA, which must be repaired by a two-protein complex known as RexAB; otherwise, the bacterium is killed, and it cannot sustain infection. This provides information on the type of damage that neutrophils cause S. aureus and the mechanism by which this damage is repaired, enabling infection.Kam Pou HaRebecca S. ClarkeGyu-Lee KimJane L. BrittanJessica E. RowleyDespoina A. I. MavridouDane ParkerThomas B. ClarkeAngela H. NobbsAndrew M. EdwardsAmerican Society for Microbiologyarticlerespiratory burstoxidative burstDNA damageEnterococcusSOS systemStaphylococcusMicrobiologyQR1-502ENmBio, Vol 11, Iss 6 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	respiratory burst oxidative burst DNA damage Enterococcus SOS system Staphylococcus Microbiology QR1-502
spellingShingle	respiratory burst oxidative burst DNA damage Enterococcus SOS system Staphylococcus Microbiology QR1-502 Kam Pou Ha Rebecca S. Clarke Gyu-Lee Kim Jane L. Brittan Jessica E. Rowley Despoina A. I. Mavridou Dane Parker Thomas B. Clarke Angela H. Nobbs Andrew M. Edwards Staphylococcal DNA Repair Is Required for Infection
description	ABSTRACT To cause infection, Staphylococcus aureus must withstand damage caused by host immune defenses. However, the mechanisms by which staphylococcal DNA is damaged and repaired during infection are poorly understood. Using a panel of transposon mutants, we identified the rexBA operon as being important for the survival of Staphylococcus aureus in whole human blood. Mutants lacking rexB were also attenuated for virulence in murine models of both systemic and skin infections. We then demonstrated that RexAB is a member of the AddAB family of helicase/nuclease complexes responsible for initiating the repair of DNA double-strand breaks. Using a fluorescent reporter system, we were able to show that neutrophils cause staphylococcal DNA double-strand breaks through reactive oxygen species (ROS) generated by the respiratory burst, which are repaired by RexAB, leading to the induction of the mutagenic SOS response. We found that RexAB homologues in Enterococcus faecalis and Streptococcus gordonii also promoted the survival of these pathogens in human blood, suggesting that DNA double-strand break repair is required for Gram-positive bacteria to survive in host tissues. Together, these data demonstrate that DNA is a target of host immune cells, leading to double-strand breaks, and that the repair of this damage by an AddAB-family enzyme enables the survival of Gram-positive pathogens during infection. IMPORTANCE To cause infection, bacteria must survive attack by the host immune system. For many bacteria, including the major human pathogen Staphylococcus aureus, the greatest threat is posed by neutrophils. These immune cells ingest the invading organisms and try to kill them with a cocktail of chemicals that includes reactive oxygen species (ROS). The ability of S. aureus to survive this attack is crucial for the progression of infection. However, it was not clear how the ROS damaged S. aureus and how the bacterium repaired this damage. In this work, we show that ROS cause breaks in the staphylococcal DNA, which must be repaired by a two-protein complex known as RexAB; otherwise, the bacterium is killed, and it cannot sustain infection. This provides information on the type of damage that neutrophils cause S. aureus and the mechanism by which this damage is repaired, enabling infection.
format	article
author	Kam Pou Ha Rebecca S. Clarke Gyu-Lee Kim Jane L. Brittan Jessica E. Rowley Despoina A. I. Mavridou Dane Parker Thomas B. Clarke Angela H. Nobbs Andrew M. Edwards
author_facet	Kam Pou Ha Rebecca S. Clarke Gyu-Lee Kim Jane L. Brittan Jessica E. Rowley Despoina A. I. Mavridou Dane Parker Thomas B. Clarke Angela H. Nobbs Andrew M. Edwards
author_sort	Kam Pou Ha
title	Staphylococcal DNA Repair Is Required for Infection
title_short	Staphylococcal DNA Repair Is Required for Infection
title_full	Staphylococcal DNA Repair Is Required for Infection
title_fullStr	Staphylococcal DNA Repair Is Required for Infection
title_full_unstemmed	Staphylococcal DNA Repair Is Required for Infection
title_sort	staphylococcal dna repair is required for infection
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/c366ab45cdbd41dbacadbfc7679c2b74
work_keys_str_mv	AT kampouha staphylococcaldnarepairisrequiredforinfection AT rebeccasclarke staphylococcaldnarepairisrequiredforinfection AT gyuleekim staphylococcaldnarepairisrequiredforinfection AT janelbrittan staphylococcaldnarepairisrequiredforinfection AT jessicaerowley staphylococcaldnarepairisrequiredforinfection AT despoinaaimavridou staphylococcaldnarepairisrequiredforinfection AT daneparker staphylococcaldnarepairisrequiredforinfection AT thomasbclarke staphylococcaldnarepairisrequiredforinfection AT angelahnobbs staphylococcaldnarepairisrequiredforinfection AT andrewmedwards staphylococcaldnarepairisrequiredforinfection
_version_	1718427174915014656

Staphylococcal DNA Repair Is Required for Infection

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