Inhibiting the stringent response blocks
            <i>Mycobacterium tuberculosis</i>
            entry into quiescence and reduces persistence

Dutta, Noton K.; Klinkenberg, Lee G.; Vázquez, María J.; Segura-Carro, Delfina; Colmenarejo, Gonzalo; Ramón, Fernando; Rodriguez-Miquel, Beatriz; Mata-Cantero, Lydia; Francisco, Esther; Chuang, Yu Min; Rubin, Harvey; Lee, Jae Jin; Eoh, Hyungjin; Bader, Joel S.; Pérez-Herrán, Esther; Mendoza-Losana, Alfonso; Karakousis, Petros C.

doi:10.1126/sciadv.aav2104

Cited by 110 publications

(133 citation statements)

References 59 publications

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“…(p)ppGpp has recently been shown to bind to Era from Staphylococcus aureus and E. coli to inhibit its GTPase activity ( 39–41 ), and this may influence ribosome biogenesis and integrity. There is good evidence that (p)ppGpp is important for development of antibiotic persisters ( 42 ) and for long term survival of latent and growth-arrested M. tuberculosis ( 43, 44 ).…”

Section: Discussionmentioning

confidence: 99%

Bacterial longevity requires protein synthesis and a stringent response

Yin

Nakayasu

et al. 2019

Preprint

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Gram-negative bacteria in infections, biofilms and industrial settings often stop growing due to nutrient depletion, immune responses or environmental stresses. Bacteria in this state tend to be tolerant to antibiotics and are often referred to as dormant. Rhodopseudomonas palustris, a phototrophic α-proteobacterium, can remain fully viable for more than four months when growth is arrested. Here, we show that protein synthesis, specific proteins involved in translation and a stringent response are required for this remarkable longevity. Because it can generate ATP from light during growth arrest, R. palustris is an extreme example of a bacterial species that will stay alive for long periods of time as a relatively homogeneous population of cells and it is thus an excellent model organism for studies of bacterial longevity. There is evidence that other Gram-negative species also continue to synthesize proteins during growth arrest and that a stringent response is required for their longevity as well. Our observations challenge the notion that growth-arrested cells are necessarily dormant and metabolically inactive, and suggest that such bacteria may have a level of metabolic activity that is higher than many would have assumed. Our results also expand our mechanistic understanding of a crucial but understudied phase of the bacterial life cycle.IMPORTANCEWe are surrounded by bacteria; but they do not completely dominate our planet despite the ability of many to grow extremely rapidly in the laboratory. This has been interpreted to mean that bacteria in nature are often in a dormant state. We investigated life in growth arrest of Rhodopseudomonas palustris, a proteobacterium that stays alive for months when it is not growing. We found that cells were metabolically active; they continued to synthesize proteins and mounted a stringent response, both of which were required for their longevity. Our results suggest that long-lived bacteria are not necessarily inactive but have an active metabolism that is well adjusted to life without growth.

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Section: Discussionmentioning

confidence: 99%

Bacterial longevity requires protein synthesis and a stringent response

Yin

Nakayasu

et al. 2019

Preprint

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“…This is supported by the finding that a M. tuberculosis mutant missing PPX1 is associated with elevated intracellular poly P and tolerance to INH ( Thayil et al, 2011 ), whereas a mutant devoid of PPK1 is associated to low levels of poly P and increased drug susceptibility ( Singh et al, 2013b ). In a recent work, Dutta et al (2019) demonstrated that a M. tuberculosis rel mtb null mutant was unable to accumulate poly P and down-modulate metabolism to decrease growth rate in a condition of nutrient starvation. While the INH minimal bactericidal concentration (MBC) of the mutant strain and its parental strain were equal in rich media, during nutrient starvation the wild type strain MBC increased 512-fold, whereas that of the mutant strain remained equal, suggesting that the stringent response activation was responsible for tolerance to this drug.…”

Section: Role Of Stringent Response In Tolerance To Drugsmentioning

confidence: 99%

“…While the INH minimal bactericidal concentration (MBC) of the mutant strain and its parental strain were equal in rich media, during nutrient starvation the wild type strain MBC increased 512-fold, whereas that of the mutant strain remained equal, suggesting that the stringent response activation was responsible for tolerance to this drug. Interestingly, enhanced susceptibility to INH of the rel mtb null mutant was also demonstrated during the chronic phase of infection in BALB/c mouse lungs ( Dutta et al, 2019 ). The finding that (p)ppGpp-mediated tolerance to INH was detectable only upon nutrient starvation, well explains the contrasting data obtained by Bhaskar et al (2018) , who did not show any tolerance to INH in different M. smegmatis mutants missing rel mtb , lon , ppk1 , ppk2 , or both ppx1 and ppx2 , as these authors run their experiments in cultures growing exponentially in rich medium, a condition not activating the stringent response.…”

Section: Role Of Stringent Response In Tolerance To Drugsmentioning

confidence: 99%

Tolerance and Persistence to Drugs: A Main Challenge in the Fight Against Mycobacterium tuberculosis

et al. 2020

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The treatment of tuberculosis is extremely long. One of the reasons why Mycobacterium tuberculosis elimination from the organism takes so long is that in particular environmental conditions it can become tolerant to drugs and/or develop persisters able to survive killing even from very high drug concentrations. Tolerance develops in response to a harsh environment exposure encountered by bacteria during infection, mainly due to the action of the immune system, whereas persistence results from the presence of heterogeneous bacterial populations with different degrees of drug sensitivity, and can be induced by exposure to stress conditions. Here, we review the actual knowledge on the stress response mechanisms enacted by M. tuberculosis during infection, which leads to increased drug tolerance or development of a highly drug-resistant subpopulation.

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“…Arresting the (p)ppGpp synthetase activity using (p)ppGpp analogues is emerging as a clinically important method in eradicating persistent infections ( de la Fuente-Núñez et al, 2014 ; Andresen et al, 2016 ; Petchiappan and Chatterji, 2017 ; Syal et al, 2017 ; Dutta et al, 2019 ). Similarly, the ssRNA-binding property of the SAS can be explored to regulate the SAS-mediated (p)ppGpp synthesis.…”

Section: Sas Bind To Ssrnamentioning

confidence: 99%

Pleiotropic Effects of Bacterial Small Alarmone Synthetases: Underscoring the Dual-Domain Small Alarmone Synthetases in Mycobacterium smegmatis

Krishnan

Chatterji

2020

Front. Microbiol.

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The nucleotide alarmone (p)ppGpp, signaling the stringent response, is known for more than 5 decades. The cellular turnover of the alarmone is regulated by RelA/SpoT homolog (RSH) superfamily of enzymes. There are long RSHs (RelA, SpoT, and Rel) and short RSHs [small alarmone synthetases (SAS) and small alarmone hydrolases (SAH)]. Long RSHs are multidomain proteins with (p)ppGpp synthesis, hydrolysis, and regulatory functions. Short RSHs are single-domain proteins with a single (p)ppGpp synthesis/hydrolysis function with few exceptions having two domains. Mycobacterial RelZ is a dual-domain SAS with RNase HII and the (p)ppGpp synthetase activity. SAS is known to impact multiple cellular functions independently and in accordance with the long RSH. Few SAS in bacteria including RelZ synthesize pGpp, the third small alarmone, along with the conventional (p)ppGpp. SAS can act as an RNA-binding protein for the negative allosteric inhibition of (p)ppGpp synthesis. Here, we initially recap the important features and molecular functions of different SAS that are previously characterized to understand the obligation for the “alarmone pool” produced by the long and short RSHs. Then, we focus on the RelZ, especially the combined functions of RNase HII and (p)ppGpp synthesis from a single polypeptide to connect with the recent findings of SAS as an RNA-binding protein. Finally, we conclude with the possibilities of using single-stranded RNA (ssRNA) as an additional therapeutic strategy to combat the persistent infections by inhibiting the redundant (p)ppGpp synthetases.

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Inhibiting the stringent response blocks Mycobacterium tuberculosis entry into quiescence and reduces persistence

Cited by 110 publications

References 59 publications

Bacterial longevity requires protein synthesis and a stringent response

Bacterial longevity requires protein synthesis and a stringent response

Tolerance and Persistence to Drugs: A Main Challenge in the Fight Against Mycobacterium tuberculosis

Pleiotropic Effects of Bacterial Small Alarmone Synthetases: Underscoring the Dual-Domain Small Alarmone Synthetases in Mycobacterium smegmatis

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