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Mycobacterium tuberculosis expresses a large number of leaderless mRNA transcripts; these lack the 5′ leader region, which usually contains the Shine–Dalgarno sequence required for translation initiation in bacteria. In M. tuberculosis , transcripts encoding proteins like toxin–antitoxin systems are predominantly leaderless and the overall ratio of leaderless to Shine–Dalgarno transcripts significantly increases during growth arrest, suggesting that leaderless translation might be important during persistence in the host. However, whether these two types of transcripts are translated with differing efficiencies during optimal growth conditions and during stress conditions that induce growth arrest, is unclear. Here, we have used the desA1 (Rv0824c) and desA2 (Rv1094) gene pair as representative for Shine–Dalgarno and leaderless transcripts in M. tuberculosis respectively; and used them to construct bioluminescent reporter strains. We detect robust leaderless translation during exponential in vitro growth, and we show that leaderless translation is more stable than Shine–Dalgarno translation during adaptation to stress conditions. These changes are independent from transcription, as transcription levels did not significantly change following quantitative real-time PCR analysis. Upon entrance into nutrient starvation and after nitric oxide exposure, leaderless translation is significantly less affected by the stress than Shine–Dalgarno translation. Similarly, during the early stages of infection of macrophages, the levels of leaderless translation are transiently more stable than those of Shine–Dalgarno translation. These results suggest that leaderless translation may offer an advantage in the physiology of M. tuberculosis. Identification of the molecular mechanisms underlying this translational regulation may provide insights into persistent infection. [ABSTRACT FROM AUTHOR]

Mycobacterium tuberculosis expresses a large number of leaderless mRNA transcripts; these lack the 5′ leader region, which usually contains the Shine–Dalgarno sequence required for translation initiation in bacteria. In M. tuberculosis, transcripts encoding proteins like toxin–antitoxin systems are predominantly leaderless and the overall ratio of leaderless to Shine–Dalgarno transcripts significantly increases during growth arrest, suggesting that leaderless translation might be important during persistence in the host. However, whether these two types of transcripts are translated with differing efficiencies during optimal growth conditions and during stress conditions that induce growth arrest, is unclear. Here, we have used the desA1 (Rv0824c) and desA2 (Rv1094) gene pair as representative for Shine–Dalgarno and leaderless transcripts in M. tuberculosis respectively; and used them to construct bioluminescent reporter strains. We detect robust leaderless translation during exponential in vitro growth, and we show that leaderless translation is more stable than Shine–Dalgarno translation during adaptation to stress conditions. These changes are independent from transcription, as transcription levels did not significantly change following quantitative real-time PCR analysis. Upon entrance into nutrient starvation and after nitric oxide exposure, leaderless translation is significantly less affected by the stress than Shine–Dalgarno translation. Similarly, during the early stages of infection of macrophages, the levels of leaderless translation are transiently more stable than those of Shine–Dalgarno translation. These results suggest that leaderless translation may offer an advantage in the physiology of M. tuberculosis. Identification of the molecular mechanisms underlying this translational regulation may provide insights into persistent infection.

Finally, chromosome segregation constitutes an essential stage of cell cycle progression, and ParA and ParB are known as main players in cellular positioning of the replication origin prior to cell division in most bacterial species. Keywords: bacterial chromosomes; nucleoid architecture; DNA replication; chromosome segregation; DNA repair; stress conditions; environmental conditions; replication-transcription conflicts EN bacterial chromosomes nucleoid architecture DNA replication chromosome segregation DNA repair stress conditions environmental conditions replication-transcription conflicts N.PAG N.PAG 3 03/16/21 20210311 NES 210311 The bacterial cell cycle comprises chromosome replication and segregation of newly replicated chromosomes into daughter cells prior to cell division. Bacterial chromosomes, nucleoid architecture, DNA replication, chromosome segregation, DNA repair, environmental conditions, replication-transcription conflicts, stress conditions In this context, Sinha et al. review the current understanding of molecular mechanisms underlying the negative impact of the stringent stress response regulator, (p)ppGpp, on chromosome replication initiation in I Escherichia coli i , and on chromosome replication elongation in I Bacillus subtilis i , respectively. [Extracted from the article]

Lycium barbarum is a boxthorn that produces the goji berries. The aim of the current study was to evaluate the proliferative effect of L. barbarum polysaccharides (LBP) on probiotics. LBP was extracted from goji berries and its monosaccharide composition characterized by gas chromatography (GC). The LBP extract contained arabinose, rhamnose, xylose, mannose, galactose, and glucose. LBP obviously promoted the proliferation of lactic acid bacteria (LAB) strains, especially Bifidobacterium longum subsp. infantis Bi-26 and Lactobacillus acidophilus NCFM. In the presence of LBP in the growth medium, the β-galactosidase (β-GAL) and lactate dehydrogenase (LDH) activities of strain Bi-26 significantly increased. The activities of β-GAL, LDH, hexokinase (HK), 6-phosphofructokinase (PFK), and pyruvate kinase (PK) of strain NCFM significantly increased under those conditions. LAB transcriptome sequencing analysis was performed to elucidate the mechanism responsible for the proliferative effect of LBP. The data revealed that LBP promoted the bacterial biosynthetic and metabolic processes, gene expression, transcription, and transmembrane transport. Pyruvate metabolism, carbon metabolism, phosphotransferase system (PTS), and glycolysis/gluconeogenesis genes were overexpressed. Furthermore, LBP improved cell vitality during freeze-drying and tolerance of the gastrointestinal environment. In summary, LBP can be used as a potential prebiotic for Bifidobacterium and Lactobacillus.

Lycium barbarum is a boxthorn that produces the goji berries. The aim of the current study was to evaluate the proliferative effect of L. barbarum polysaccharides (LBP) on probiotics. LBP was extracted from goji berries and its monosaccharide composition characterized by gas chromatography (GC). The LBP extract contained arabinose, rhamnose, xylose, mannose, galactose, and glucose. LBP obviously promoted the proliferation of lactic acid bacteria (LAB) strains, especially Bifidobacterium longum subsp. infantis Bi-26 and Lactobacillus acidophilus NCFM. In the presence of LBP in the growth medium, the b-galactosidase (b-GAL) and lactate dehydrogenase (LDH) activities of strain Bi-26 significantly increased. The activities of b-GAL, LDH, hexokinase (HK), 6-phosphofructokinase (PFK), and pyruvate kinase (PK) of strain NCFM significantly increased under those conditions. LAB transcriptome sequencing analysis was performed to elucidate the mechanism responsible for the proliferative effect of LBP. The data revealed that LBP promoted the bacterial biosynthetic and metabolic processes, gene expression, transcription, and transmembrane transport. Pyruvate metabolism, carbon metabolism, phosphotransferase system (PTS), and glycolysis/gluconeogenesis genes were overexpressed. Furthermore, LBP improved cell vitality during freeze-drying and tolerance of the gastrointestinal environment. In summary, LBP can be used as a potential prebiotic for Bifidobacterium and Lactobacillus. [ABSTRACT FROM AUTHOR]

Pathogenic bacterial persisters are responsible for the recalcitrance of chronic and persistent infections to antimicrobial therapy. Although the mechanisms of persister formation and survival have been widely studied in Escherichia coli, persistence mechanisms in S. aureus remain largely unknown. Here, we screened a transposon mutant library of a clinical methicillin-resistant Staphylococcus aureus（MRSA）strain, USA500 (ST8), under antibiotic pressure and identified 13 genes whose insertion mutations resulted in a defect in persistence. These candidate genes were further confirmed by evaluating the survival of the mutants upon exposure to levofloxacin and several other stress conditions. We found 13 insertion mutants with significantly lower persister numbers under several stress conditions, including sdhA, sdhB, ureG, mnhG1, fbaA, ctaB, clpX, parE, HOU_0223, HOU_0587, HOU_2091, HOU_2315 and HOU_2346, which mapped into pathways of oxidative phosphorylation, TCA cycle, glycolysis, cell cycle and ABC transporters, suggesting that these genes and pathways may play an important role in persister formation and survival. The newly constructed knockout strains of ureG, sdhA and sdhB and their complemented strains were also tested for defect in persisters following exposure to levofloxacin and several other stress conditions. The results from these experiments were consistent with the screening results, which indicated that deletion of these genes in MRSA USA500 leads to persister defect. These findings provide novel insights into the mechanisms of persister formation and survival in S. aureus and offer new targets for the development of persister-directed antibiotics for the improved treatment of chronic and persistent infections.

Lycium barbarum is a boxthorn that produces the goji berries. The aim of the current study was to evaluate the proliferative effect of L. barbarum polysaccharides (LBP) on probiotics. LBP was extracted from goji berries and its monosaccharide composition characterized by gas chromatography (GC). The LBP extract contained arabinose, rhamnose, xylose, mannose, galactose, and glucose. LBP obviously promoted the proliferation of lactic acid bacteria (LAB) strains, especially Bifidobacterium longum subsp. infantis Bi-26 and Lactobacillus acidophilus NCFM. In the presence of LBP in the growth medium, the β-galactosidase (β-GAL) and lactate dehydrogenase (LDH) activities of strain Bi-26 significantly increased. The activities of β-GAL, LDH, hexokinase (HK), 6-phosphofructokinase (PFK), and pyruvate kinase (PK) of strain NCFM significantly increased under those conditions. LAB transcriptome sequencing analysis was performed to elucidate the mechanism responsible for the proliferative effect of LBP. The data revealed that LBP promoted the bacterial biosynthetic and metabolic processes, gene expression, transcription, and transmembrane transport. Pyruvate metabolism, carbon metabolism, phosphotransferase system (PTS), and glycolysis/gluconeogenesis genes were overexpressed. Furthermore, LBP improved cell vitality during freeze-drying and tolerance of the gastrointestinal environment. In summary, LBP can be used as a potential prebiotic for Bifidobacterium and Lactobacillus .