Journal of Bacteriology最新文献

{"title":"The metal-binding GTPases CobW2 and CobW3 are at the crossroads of zinc and cobalt homeostasis in <i>Cupriavidus metallidurans</i>.","authors":"Diana Galea, Martin Herzberg, Dietrich H Nies","doi":"10.1128/jb.00226-24","DOIUrl":"10.1128/jb.00226-24","url":null,"abstract":"<p><p>The metal-resistant beta-proteobacterium <i>Cupriavidus metallidurans</i> is also able to survive conditions of metal starvation. We show that zinc-starved cells can substitute some of the required zinc with cobalt but not with nickel ions. The zinc importer ZupT was necessary for this process but was not essential for either zinc or cobalt import. The cellular cobalt content was also influenced by the two COG0523-family proteins, CobW2 and CobW3. Pulse-chase experiments with radioactive and isotope-enriched zinc demonstrated that both proteins interacted with ZupT to control the cellular flow-equilibrium of zinc, a central process of zinc homeostasis. Moreover, an antagonistic interplay of CobW2 and CobW3 in the presence of added cobalt caused a growth defect in mutant cells devoid of the cobalt efflux system DmeF. Full cobalt resistance also required a synergistic interaction of ZupT and DmeF. Thus, the two transporters along with CobW2 and CobW3 interact to control cobalt homeostasis in a process that depends on zinc availability. Because ZupT, CobW2, and CobW3 also direct zinc homeostasis, this process links the control of cobalt and zinc homeostasis, which subsequently protects <i>C. metallidurans</i> against cadmium stress and general metal starvation.IMPORTANCEIn bacterial cells, zinc ions need to be allocated to zinc-dependent proteins without disturbance of this process by other transition metal cations. Under zinc-starvation conditions, <i>C. metallidurans</i> floods the cell with cobalt ions, which protect the cell against cadmium toxicity, help withstand metal starvation, and provide cobalt to metal-promiscuous paralogs of essential zinc-dependent proteins. The number of cobalt ions needs to be carefully controlled to avoid a toxic cobalt overload. This is accomplished by an interplay of the zinc importer ZupT with the COG0523-family proteins, CobW3, and CobW2. At high external cobalt concentrations, this trio of proteins additionally interacts with the cobalt efflux system, DmeF, so that these four proteins form an inextricable link between zinc and cobalt homeostasis.</p>","PeriodicalId":15107,"journal":{"name":"Journal of Bacteriology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11340326/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141748301","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>Salmonella</i> Typhimurium alters galactitol metabolism under ciprofloxacin treatment to balance resistance and virulence.","authors":"Qiwei Chen, Yongfeng Yu, Yongchang Xu, Heng Quan, Donghui Liu, Caiyu Li, Mengyao Liu, Xiaowei Gong","doi":"10.1128/jb.00178-24","DOIUrl":"10.1128/jb.00178-24","url":null,"abstract":"<p><p>Ciprofloxacin-resistant <i>Salmonella</i> Typhimurium (<i>S.</i> Typhimurium) causes a significant health burden worldwide. A wealth of studies has been published on the contributions of different mechanisms to ciprofloxacin resistance in <i>Salmonella</i> spp. But we still lack a deep understanding of the physiological responses and genetic changes that underlie ciprofloxacin exposure. This study aims to know how phenotypic and genotypic characteristics are impacted by ciprofloxacin exposure, from ciprofloxacin-susceptible to ciprofloxacin-resistant strains <i>in vitro</i>. Here, we investigated the multistep evolution of resistance in replicate populations of <i>S.</i> Typhimurium during 24 days of continuously increasing ciprofloxacin exposure and assessed how ciprofloxacin impacts physiology and genetics. Numerous studies have demonstrated that RamA is a global transcriptional regulator that prominently perturbs the transcriptional landscape of <i>S.</i> Typhimurium, resulting in a ciprofloxacin-resistant phenotype appearing first; the quinolone resistance-determining region mutation site can only be detected later. Comparing the microbial physiological changes and RNA sequencing (RNA-Seq) results of ancestral and selectable mutant strains, the selectable mutant strains had some fitness costs, such as decreased virulence, an increase of biofilm-forming ability, a change of \"collateral\" sensitivity to other drugs, and inability to utilize galactitol. Importantly, in the ciprofloxacin induced, RamA directly binds and activates the <i>gatR</i> gene responsible for the utilization of galactitol, but RamA deletion strains could not activate <i>gatR</i>. The elevated levels of RamA, which inhibit the galactitol metabolic pathway through the activation of <i>gatR</i>, can lead to a reduction in the growth rate, adhesion, and colonization resistance of <i>S</i>. Typhimurium. This finding is supported by studies conducted in M9 medium as well as <i>in vivo</i> infection models.</p><p><strong>Importance: </strong>Treatment of antibiotic resistance can significantly benefit from a deeper understanding of the interactions between drugs and genetics. The physiological responses and genetic mechanisms in antibiotic-exposed bacteria are not well understood. Traditional resistance studies, often retrospective, fail to capture the entire resistance development process and typically exhibit unpredictable dynamics. To explore how clinical isolates of <i>S.</i> Typhimurium respond to ciprofloxacin, we analyzed their adaptive responses. We found that <i>S.</i> Typhimurium RamA-mediated regulation disrupts microbial metabolism under ciprofloxacin exposure, affecting genes in the galactitol metabolic pathways. This disruption facilitates adaptive responses to drug therapy and enhances the efficiency of intracellular survival. A more comprehensive and integrated understanding of these physiological and genetic changes is crucial for improving treatment outcome","PeriodicalId":15107,"journal":{"name":"Journal of Bacteriology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11340313/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141855629","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcriptional and post-transcriptional mechanisms modulate cyclopropane fatty acid synthase through small RNAs in <i>Escherichia coli</i>.","authors":"Colleen M Bianco, Nancy N Caballero-Rothar, Xiangqian Ma, Kristen R Farley, Carin K Vanderpool","doi":"10.1128/jb.00049-24","DOIUrl":"10.1128/jb.00049-24","url":null,"abstract":"<p><p>The small RNA (sRNA) RydC strongly activates <i>cfa</i>, which encodes the cyclopropane fatty acid synthase. Previous work demonstrated that RydC activation of <i>cfa</i> increases the conversion of unsaturated fatty acids to cyclopropanated fatty acids in membrane lipids and changes the biophysical properties of membranes, making cells more resistant to acid stress. The regulators that control RydC synthesis had not previously been identified. In this study, we identify a GntR-family transcription factor, YieP, that represses <i>rydC</i> transcription. YieP positively autoregulates its own transcription and indirectly regulates <i>cfa</i> through RydC. We further identify additional sRNA regulatory inputs that contribute to the control of RydC and <i>cfa</i>. The translation of <i>yieP</i> is repressed by the Fnr-dependent sRNA, FnrS, making FnrS an indirect activator of <i>rydC</i> and <i>cfa</i>. Conversely, RydC activity on <i>cfa</i> is antagonized by the OmpR-dependent sRNA OmrB. Altogether, this work illuminates a complex regulatory network involving transcriptional and post-transcriptional inputs that link the control of membrane biophysical properties to multiple environmental signals.</p><p><strong>Importance: </strong>Bacteria experience many environmental stresses that challenge their membrane integrity. To withstand these challenges, bacteria sense what stress is occurring and mount a response that protects membranes. Previous work documented the important roles of small RNA (sRNA) regulators in membrane stress responses. One sRNA, RydC, helps cells cope with membrane-disrupting stresses by promoting changes in the types of lipids incorporated into membranes. In this study, we identified a regulator, YieP, that controls when RydC is produced and additional sRNA regulators that modulate YieP levels and RydC activity. These findings illuminate a complex regulatory network that helps bacteria sense and respond to membrane stress.</p>","PeriodicalId":15107,"journal":{"name":"Journal of Bacteriology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11340327/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141558829","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The role of metals in hypothiocyanite resistance in <i>Escherichia coli</i>.","authors":"Michael J Gray","doi":"10.1128/jb.00098-24","DOIUrl":"10.1128/jb.00098-24","url":null,"abstract":"<p><p>The innate immune system employs a variety of antimicrobial oxidants to control and kill host-associated bacteria. Hypothiocyanite/hypothiocyanous acid (^-OSCN/HOSCN) is one such antimicrobial oxidant that is synthesized by lactoperoxidase, myeloperoxidase, and eosinophil peroxidase at sites throughout the human body. HOSCN has potent antibacterial activity while being largely non-toxic toward human cells. The molecular mechanisms by which bacteria sense and defend themselves against HOSCN have only recently begun to be elaborated, notably by the discovery of bacterial HOSCN reductase (RclA), an HOSCN-degrading enzyme widely conserved among bacteria that live on epithelial surfaces. In this paper, I show that Ni²⁺ sensitizes <i>Escherichia coli</i> to HOSCN by inhibiting glutathione reductase and that inorganic polyphosphate protects <i>E. coli</i> against this effect, probably by chelating Ni²⁺ ions. I also found that RclA is very sensitive to inhibition by Cu²⁺ and Zn²⁺, metals that are accumulated to high levels by innate immune cells, and that, surprisingly, thioredoxin and thioredoxin reductase are not involved in HOSCN stress resistance in <i>E. coli</i>. These results advance our understanding of the contribution of different oxidative stress responses and redox buffering pathways to HOSCN resistance in <i>E. coli</i> and illustrate important interactions between metal ions and the enzymes bacteria use to defend themselves against oxidative stress.</p><p><strong>Importance: </strong>Hypothiocyanite (HOSCN) is an antimicrobial oxidant produced by the innate immune system. The molecular mechanisms by which host-associated bacteria defend themselves against HOSCN have only recently begun to be understood. The results in this paper are significant because they show that the low molecular weight thiol glutathione and enzyme glutathione reductase are critical components of the <i>Escherichia coli</i> HOSCN response, working by a mechanism distinct from that of the HOSCN-specific defenses provided by the RclA, RclB, and RclC proteins and that metal ions (including nickel, copper, and zinc) may impact the ability of bacteria to resist HOSCN by inhibiting specific defensive enzymes (e.g., glutathione reductase or RclA).</p>","PeriodicalId":15107,"journal":{"name":"Journal of Bacteriology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11340325/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141626734","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dissecting structure and function of the monovalent cation/H⁺ antiporters Mdm38 and Ylh47 in <i>Saccharomyces cerevisiae</i>.","authors":"Masaru Tsujii, Ellen Tanudjaja, Haoyu Zhang, Haruto Shimizukawa, Ayumi Konishi, Tadaomi Furuta, Yasuhiro Ishimaru, Nobuyuki Uozumi","doi":"10.1128/jb.00182-24","DOIUrl":"10.1128/jb.00182-24","url":null,"abstract":"<p><p><i>Saccharomyces cerevisiae</i> Mdm38 and Ylh47 are homologs of the Ca²⁺/H⁺ antiporter Letm1, a candidate gene for seizures associated with Wolf-Hirschhorn syndrome in humans. Mdm38 is important for K⁺/H⁺ exchange across the inner mitochondrial membrane and contributes to membrane potential formation and mitochondrial protein translation. Ylh47 also localizes to the inner mitochondrial membrane. However, knowledge of the structures and detailed transport activities of Mdm38 and Ylh47 is limited. In this study, we conducted characterization of the ion transport activities and related structural properties of Mdm38 and Ylh47. Growth tests using Na⁺/H⁺ antiporter-deficient <i>Escherichia coli</i> strain TO114 showed that Mdm38 and Ylh47 had Na⁺ efflux activity. Measurement of transport activity across <i>E. coli</i>-inverted membranes showed that Mdm38 and Ylh47 had K⁺/H⁺, Na⁺/H⁺, and Li⁺/H⁺ antiport activity, but unlike Letm1, they lacked Ca²⁺/H⁺ antiport activity. Deletion of the ribosome-binding domain resulted in decreased Na⁺ efflux activity in Mdm38. Structural models of Mdm38 and Ylh47 identified a highly conserved glutamic acid in the pore-forming membrane-spanning region. Replacement of this glutamic acid with alanine, a non-polar amino acid, significantly impaired the ability of Mdm38 and Ylh47 to complement the salt sensitivity of <i>E. coli</i> TO114. These findings not only provide important insights into the structure and function of the Letm1-Mdm38-Ylh47 antiporter family but by revealing their distinctive properties also shed light on the physiological roles of these transporters in yeast and animals.</p><p><strong>Importance: </strong>The inner membrane of mitochondria contains numerous ion transporters, including those facilitating H⁺ transport by the electron transport chain and ATP synthase to maintain membrane potential. Letm1 in the inner membrane of mitochondria in animals functions as a Ca²⁺/H⁺ antiporter. However, this study reveals that homologous antiporters in mitochondria of yeast, Mdm38 and Ylh47, do not transport Ca²⁺ but instead are selective for K⁺ and Na⁺. Additionally, the identification of conserved amino acids crucial for antiporter activity further expanded our understanding of the structure and function of the Letm1-Mdm38-Ylh47 antiporter family.</p>","PeriodicalId":15107,"journal":{"name":"Journal of Bacteriology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11340316/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141855630","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction for Gao et al., \"Identification of the Regulon of AphB and Its Essential Roles in LuxR and Exotoxin Asp Expression in the Pathogen <i>Vibrio alginolyticus</i>\".","authors":"Xiating Gao, Yang Liu, Huan Liu, Zheng Yang, Qin Liu, Yuanxing Zhang, Qiyao Wang","doi":"10.1128/jb.00192-24","DOIUrl":"10.1128/jb.00192-24","url":null,"abstract":"","PeriodicalId":15107,"journal":{"name":"Journal of Bacteriology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11340298/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141616487","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CRP improves the survival and competitive fitness of <i>Salmonella</i> Typhimurium under starvation by controlling the cellular maintenance rate.","authors":"L K Mishra, R Shashidhar","doi":"10.1128/jb.00010-24","DOIUrl":"10.1128/jb.00010-24","url":null,"abstract":"<p><p>Catabolite repression is a mechanism of selectively utilizing preferred nutrient sources by redirecting the metabolic pathways. Therefore, it prevents non-essential energy expenditure by repressing the genes and proteins involved in the metabolism of other less favored nutrient sources. Catabolite repressor protein (CRP) is a chief mediator of catabolite repression in microorganisms. In this context, we investigated the role of CRP in starvation tolerance, at both cell physiology and molecular level, by comparing the growth, survival, competitive fitness, maintenance rate, and gene and protein expression of wild type (WT) and ∆<i>crp</i> of <i>Salmonella</i> Typhimurium, under nutrient-rich and minimal medium condition. The ∆<i>crp</i> shows slow growth upon the arrival of nutrient-limiting conditions, poor survival under prolong-starvation, and inability to compete with its counterpart WT strain in nutrient-rich [Luria broth (LB)] and glucose-supplemented M9 minimal medium. Surprisingly, we observed that the survival and competitive fitness of ∆<i>crp</i> are influenced by the composition of the growth medium. Consequently, compared to the glucose-supplemented M9 medium, ∆<i>crp</i> shows faster death and a higher maintenance rate in the LB medium. The comparative gene and protein expression studies of WT and ∆<i>crp</i> in LB medium show that ∆<i>crp</i> has partial or complete loss of repression from CRP-controlled genes, resulting in a high abundance of hundreds of proteins in ∆<i>crp</i> compared to WT. Subsequently, the addition of metabolizable sugar or fresh nutrients to the competing culture showed extended survival of ∆<i>crp</i>. Therefore, our results suggest that CRP-mediated gene repression improves starvation tolerance and competitive fitness of <i>Salmonella</i> Typhimurium by adapting its cellular maintenance rate to environmental conditions.IMPORTANCE<i>Salmonella</i> Typhimurium is a master at adapting to chronic starvation conditions. However, the molecular mechanisms to adapt to such conditions are still unknown. In this context, we have evaluated the role of catabolite repressor protein (CRP), a dual transcriptional regulator, in providing survival and competitive fitness under starvation conditions. Also, it showed an association between CRP and nutrient composition. We observed that Δ<i>crp</i> growing on alternate carbon sources has lower survival and competitive fitness than Δ<i>crp</i> growing on glucose as a carbon source. We observed that this is due to the loss of repression from the glucose and CRP-controlled genes, resulting in elevated cellular metabolism (a high maintenance rate) of the Δ<i>crp</i> during growth in a medium having a carbon source other than glucose (e.g., Luria broth medium).</p>","PeriodicalId":15107,"journal":{"name":"Journal of Bacteriology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11340309/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141751797","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>Coxiella burnetii</i> inhibits nuclear translocation of TFEB, the master transcription factor for lysosomal biogenesis.","authors":"Brigham Killips, Emily J Bremer Heaton, Leonardo Augusto, Anders Omsland, Stacey D Gilk","doi":"10.1128/jb.00150-24","DOIUrl":"10.1128/jb.00150-24","url":null,"abstract":"<p><p><i>Coxiella burnetii</i> is a highly infectious, Gram-negative, obligate intracellular bacterium and the causative agent of human Q fever. The <i>Coxiella</i> Containing Vacuole (CCV) is a modified phagolysosome that forms through fusion with host endosomes and lysosomes. While an initial acidic pH < 4.7 is essential to activate <i>Coxiella</i> metabolism, the mature, growth-permissive CCV has a luminal pH of ~5.2 that remains stable throughout infection. Inducing CCV acidification to a lysosomal pH (~4.7) causes <i>Coxiella</i> degradation, suggesting that <i>Coxiella</i> regulates CCV pH. Supporting this hypothesis, <i>Coxiella</i> blocks host lysosomal biogenesis, leading to fewer host lysosomes available to fuse with the CCV. Host cell lysosome biogenesis is primarily controlled by the transcription factor EB (TFEB), which binds Coordinated Lysosomal Expression And Regulation (CLEAR) motifs upstream of genes involved in lysosomal biogenesis and function. TFEB is a member of the microphthalmia/transcription factor E (MiT/TFE) protein family, which also includes MITF, TFE3, and TFEC. This study examines the roles of MiT/TFE proteins during <i>Coxiella</i> infection. We found that in cells lacking TFEB, both <i>Coxiella</i> growth and CCV size increase. Conversely, TFEB overexpression or expression in the absence of other family members leads to significantly less bacterial growth and smaller CCVs. TFE3 and MITF do not appear to play a significant role during <i>Coxiella</i> infection. Surprisingly, we found that <i>Coxiella</i> actively blocks TFEB nuclear translocation in a Type IV Secretion System-dependent manner, thus decreasing lysosomal biogenesis. Together, these results suggest that <i>Coxiella</i> inhibits TFEB nuclear translocation to limit lysosomal biogenesis, thus avoiding further CCV acidification through CCV-lysosomal fusion.</p><p><strong>Importance: </strong>The obligate intracellular bacterial pathogen <i>Coxiella burnetii</i> causes the zoonotic disease Q fever, which is characterized by a debilitating flu-like illness in acute cases and life-threatening endocarditis in patients with chronic disease. While <i>Coxiella</i> survives in a unique lysosome-like vacuole called the <i>Coxiella</i> Containing Vacuole (CCV), the bacterium inhibits lysosome biogenesis as a mechanism to avoid increased CCV acidification. Our results establish that transcription factor EB (TFEB), a member of the microphthalmia/transcription factor E (MiT/TFE) family of transcription factors that regulate lysosomal gene expression, restricts <i>Coxiella</i> infection. Surprisingly, <i>Coxiella</i> blocks TFEB translocation from the cytoplasm to the nucleus, thus downregulating the expression of lysosomal genes. These findings reveal a novel bacterial mechanism to regulate lysosomal biogenesis.</p>","PeriodicalId":15107,"journal":{"name":"Journal of Bacteriology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11340324/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141758837","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The characteristics of autolysins associated with cell separation in <i>Bacillus subtilis</i>.","authors":"Rui Li, Ronghao Chu, Rui Ban","doi":"10.1128/jb.00133-24","DOIUrl":"10.1128/jb.00133-24","url":null,"abstract":"<p><p>The peptidoglycan hydrolases responsible for the cell separation of <i>Bacillus subtilis</i> cells are collectively referred to as autolysins. However, the role of each autolysin in the cell separation of <i>B. subtilis</i> is not fully understood. In this study, we constructed a series of cell separation-associated autolysin deficient strains and strains overexpressing the transcription factors SlrR and SinR, and the morphological changes of these strains in liquid culture were observed. The results showed that the absence of D,L-endopeptidases CwlS and LytF only increased the cell chain length in the early exponential phase. The absence of D,L-endopeptidase LytE or N-acetylmuramyl-L-alanine amidase LytC can cause cells to form chains throughout the growth of <i>B. subtilis</i>, although the cell chain length was significantly shortened during the stationary phase. However, the absence of peptidoglycan N-acetylglucosaminidase LytD only caused minor defect in cell separation. Therefore, we concluded that LytE and LytC were the major autolysins that ensure the timely separation of <i>B. subtilis</i> daughter cells, whereas CwlS, LytF, and LytD were the minor autolysins. In addition, overexpression of the transcription factors SinR and SlrR in the <i>cwlS lytF lytC lytE</i> mutant enabled <i>B. subtilis</i> cells to form ultra-long chains in the vegetative phase, and its biomass level was basically the same as that of the wild type. This led to the conclusion that besides inhibiting the expression of <i>lytC</i> and <i>lytF</i>, the SinR-SlrR complex also has other potential mechanisms to inhibit cell separation.<b>IMPORTANCE</b>In this study, the effects of CwlS, LytC, LytD, LytF, LytE, and SinR-SlrR complex on the cell separation of <i>Bacillus subtilis</i> at different growth phases were studied, and an ultra-long-chained <i>B. subtilis</i> strain was constructed. In microbial fermentation, due to its large cell size, this ultra-long-chained <i>B. subtilis</i> strain may be more likely to be precipitated or intercepted during the removal of bacterial process with centrifugation and membrane filtration as the main methods, which is crucial to improve the purity of the product.</p>","PeriodicalId":15107,"journal":{"name":"Journal of Bacteriology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11340307/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141620065","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An insight into the role of branched-chain α-keto acid dehydrogenase (BKD) complex in branched-chain fatty acid biosynthesis and virulence of <i>Listeria monocytogenes</i>.","authors":"Q M Monzur Kader Chowdhury, Shamima Islam, Lakshmi Narayanan, Seto C Ogunleye, Shangshang Wang, Dinh Thu, Nancy E Freitag, Mark L Lawrence, Hossam Abdelhamed","doi":"10.1128/jb.00033-24","DOIUrl":"10.1128/jb.00033-24","url":null,"abstract":"<p><p><i>Listeria monocytogenes</i> is a foodborne bacterial pathogen that causes listeriosis. Positive regulatory factor A (PrfA) is a pleiotropic master activator of virulence genes of <i>L. monocytogenes</i> that becomes active upon the entry of the bacterium into the cytosol of infected cells. <i>L. monocytogenes</i> can survive and multiply at low temperatures; this is accomplished through the maintenance of appropriate membrane fluidity via branched-chain fatty acid (BCFA) synthesis. Branched-chain α-keto acid dehydrogenase (BKD), which is composed of four polypeptides encoded by <i>lpd</i>, <i>bkdA1</i>, <i>bkdA2</i>, and <i>bkdB</i>, is known to play a vital role in BCFA biosynthesis. Here, we constructed BKD-deficient <i>Listeria</i> strains by in-frame deletion of <i>lpd</i>, <i>bkdA1</i>, <i>bkdA2</i>, and <i>bkdB</i> genes. To determine the role in <i>in vivo</i> and <i>in vitro</i>, mouse model challenges, plaque assay in murine L2 fibroblast, and intracellular replication in J744A.1 macrophage were conducted. BKD-deficient strains exhibited defects in BCFA composition, virulence, and PrfA-regulon function within the host cells. Transcriptomics analysis revealed that the transcript level of the PrfA-regulon was lower in Δ<i>bkdA1</i> strain than those in the wild-type. This study demonstrates that <i>L. monocytogenes</i> strains lacking BKD complex components were defective in PrfA-regulon function, and full activation of wild-type <i>prfA</i> may not occur within host cells in the absence of BKD. Further study will investigate the consequences of BKD deletion on PrfA function through altering BCFA catabolism.IMPORTANCE<i>Listeria monocytogenes</i> is the causative agent of listeriosis, a disease with a high mortality rate. In this study, we have shown that the deletion of BKD can impact the function of PrfA and the PrfA-regulon. The production of virulence proteins within host cells is necessary for <i>L. monocytogenes</i> to promote its intracellular survival and is likely dependent on membrane integrity. We thus report a link between <i>L. monocytogenes</i> membrane integrity and the function of PrfA. This knowledge will increase our understanding of <i>L. monocytogenes</i> pathogenesis, which may provide insight into the development of antimicrobial agents.</p>","PeriodicalId":15107,"journal":{"name":"Journal of Bacteriology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11270904/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141426955","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}