Pub Date : 2024-04-30eCollection Date: 2024-01-01DOI: 10.15698/mic2024.04.820
Pablo Gallardo, Mariana Izquierdo, Tomeu Viver, Esteban Bustos-Caparros, Dana Piras, Roberto M Vidal, Hermie M J Harmsen, Mauricio J Farfan
Diarrheagenic Escherichia coli (DEC) is the main cause of diarrhea in children under five years old. The virulence of DEC is tightly regulated by environmental signals influenced by the gut microbiota and its metabolites. Short-chain fatty acids (SCFAs) are the main metabolic product of anaerobic fermentation in the gut, but their role in DEC diarrhea has not yet been established. In this study, we determine the levels of acetate, propionate, and butyrate in stool samples from children with diarrhea caused by DEC, and we identify bacteria from the fecal gut microbiota associated with the production of SCFAs. The microbiota and SCFAs levels in stool samples obtained from 40 children with diarrhea and 43 healthy children were determined by 16S rRNA gene sequencing and HPLC, respectively. Additionally, shotgun metagenomics was used to identify metagenome-assembled genomes (MAGs) in a subgroup of samples. The results showed significantly higher levels of all SCFAs tested in diarrheal samples than in healthy controls. The abundance of Streptococcus sp., Limosilactobacillus, Blautia, Escherichia, Bacteroides, Megamonas, and Roseburia was higher in the DEC group than in healthy individuals. Functional analysis of bacteria and their main metabolic pathways made it possible to identify species MAGs that could be responsible for the detected SCFAs levels in DEC-positive diarrhea. In conclusion, based on our results and published data, we suggest that SCFAs may be important in the crosstalk between the microbiota and DEC pathogens in the gut.
致腹泻大肠杆菌(DEC)是导致五岁以下儿童腹泻的主要原因。DEC 的致病力受到肠道微生物群及其代谢产物影响的环境信号的严格调控。短链脂肪酸(SCFA)是肠道厌氧发酵的主要代谢产物,但其在 DEC 腹泻中的作用尚未确定。在本研究中,我们测定了由 DEC 引起的腹泻患儿粪便样本中乙酸盐、丙酸盐和丁酸盐的含量,并从粪便肠道微生物群中找出了与 SCFAs 的产生有关的细菌。通过 16S rRNA 基因测序和高效液相色谱法,分别测定了 40 名腹泻患儿和 43 名健康儿童粪便样本中的微生物群和 SCFAs 含量。此外,研究人员还利用霰弹枪元基因组学鉴定了一部分样本中的元基因组组装基因组(MAGs)。结果显示,腹泻样本中所有受测 SCFAs 的含量均明显高于健康对照组。在 DEC 组中,链球菌、Limosilactobacillus、Blautia、Escherichia、Bacteroides、Megamonas 和 Roseburia 的含量高于健康人。通过对细菌及其主要代谢途径进行功能分析,可以确定在 DEC 阳性腹泻中检测到的 SCFAs 水平可能是由 MAGs 物种引起的。总之,根据我们的研究结果和已发表的数据,我们认为 SCFAs 可能在肠道微生物群和 DEC 病原体之间的相互影响中起着重要作用。
{"title":"A metagenomic approach to unveil the association between fecal gut microbiota and short-chain fatty acids in diarrhea caused by diarrheagenic <i>Escherichia coli</i> in children.","authors":"Pablo Gallardo, Mariana Izquierdo, Tomeu Viver, Esteban Bustos-Caparros, Dana Piras, Roberto M Vidal, Hermie M J Harmsen, Mauricio J Farfan","doi":"10.15698/mic2024.04.820","DOIUrl":"10.15698/mic2024.04.820","url":null,"abstract":"<p><p>Diarrheagenic <i>Escherichia coli</i> (DEC) is the main cause of diarrhea in children under five years old. The virulence of DEC is tightly regulated by environmental signals influenced by the gut microbiota and its metabolites. Short-chain fatty acids (SCFAs) are the main metabolic product of anaerobic fermentation in the gut, but their role in DEC diarrhea has not yet been established. In this study, we determine the levels of acetate, propionate, and butyrate in stool samples from children with diarrhea caused by DEC, and we identify bacteria from the fecal gut microbiota associated with the production of SCFAs. The microbiota and SCFAs levels in stool samples obtained from 40 children with diarrhea and 43 healthy children were determined by 16S rRNA gene sequencing and HPLC, respectively. Additionally, shotgun metagenomics was used to identify metagenome-assembled genomes (MAGs) in a subgroup of samples. The results showed significantly higher levels of all SCFAs tested in diarrheal samples than in healthy controls. The abundance of <i>Streptococcus</i> sp., <i>Limosilactobacillus</i>, <i>Blautia</i>, <i>Escherichia</i>, <i>Bacteroides</i>, <i>Megamonas,</i> and <i>Roseburia</i> was higher in the DEC group than in healthy individuals. Functional analysis of bacteria and their main metabolic pathways made it possible to identify species MAGs that could be responsible for the detected SCFAs levels in DEC-positive diarrhea. In conclusion, based on our results and published data, we suggest that SCFAs may be important in the crosstalk between the microbiota and DEC pathogens in the gut.</p>","PeriodicalId":18397,"journal":{"name":"Microbial Cell","volume":"11 ","pages":"116-127"},"PeriodicalIF":4.1,"publicationDate":"2024-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11122282/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141155358","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}
Pub Date : 2024-04-04eCollection Date: 2024-01-01DOI: 10.15698/mic2024.03.819
Andreea-Cristina Paraschiv, Vitalie Vacaras, Cristina Nistor, Cristiana Vacaras, Stefan Strilciuc, Dafin F Muresanu
Gut microbiota has complex immune functions, related to different pathologies, including multiple sclerosis (MS).This study evaluated the influence of treatments on gut microbiota in people with MS (PwMS). The research comprised 60 participants, including 39 PwMS and 21 healthy controls (HC). Among the PwMS, 20 were prescribed a disease-modifying therapy (DMT), either interferon beta1a or teriflunomide, while 19 received a combination of classical DMT and an immunoglobulin Y (IgY) supplement. For each participant, two sets of gut samples were collected: one at the study's outset and another after two months. Alpha and beta diversity analyses revealed no significant differences between groups. In comparison to the HC, the MS group exhibited an increase in Prevotella stercorea and a decrease in Faecalibacterium prausnitzii. Following treatment, individuals with MS showed enrichment in Lachnospiraceae and Streptococcus. The second sample, compared to the first one, demonstrated an increase in Bifidobacterium angulatum and a decrease in Oscillospira for individuals with MS. Gut microbiota diversity in PwMS is not significantly different to HC.However, specific taxonomic changes indicate the presence of a dysbiosis state. The use of DMTs and immunoglobulin Y supplements may contribute to alterations in microbial composition, potentially leading to the restoration of a healthier microbiome.
{"title":"The effect of multiple sclerosis therapy on gut microbiota dysbiosis: a longitudinal prospective study.","authors":"Andreea-Cristina Paraschiv, Vitalie Vacaras, Cristina Nistor, Cristiana Vacaras, Stefan Strilciuc, Dafin F Muresanu","doi":"10.15698/mic2024.03.819","DOIUrl":"https://doi.org/10.15698/mic2024.03.819","url":null,"abstract":"<p><p>Gut microbiota has complex immune functions, related to different pathologies, including multiple sclerosis (MS).This study evaluated the influence of treatments on gut microbiota in people with MS (PwMS). The research comprised 60 participants, including 39 PwMS and 21 healthy controls (HC). Among the PwMS, 20 were prescribed a disease-modifying therapy (DMT), either interferon beta1a or teriflunomide, while 19 received a combination of classical DMT and an immunoglobulin Y (IgY) supplement. For each participant, two sets of gut samples were collected: one at the study's outset and another after two months. Alpha and beta diversity analyses revealed no significant differences between groups. In comparison to the HC, the MS group exhibited an increase in <i>Prevotella stercorea</i> and a decrease in <i>Faecalibacterium prausnitzii</i>. Following treatment, individuals with MS showed enrichment in <i>Lachnospiraceae</i> and <i>Streptococcus</i>. The second sample, compared to the first one, demonstrated an increase in <i>Bifidobacterium angulatum</i> and a decrease in <i>Oscillospira</i> for individuals with MS. Gut microbiota diversity in PwMS is not significantly different to HC.However, specific taxonomic changes indicate the presence of a dysbiosis state. The use of DMTs and immunoglobulin Y supplements may contribute to alterations in microbial composition, potentially leading to the restoration of a healthier microbiome.</p>","PeriodicalId":18397,"journal":{"name":"Microbial Cell","volume":"11 ","pages":"106-115"},"PeriodicalIF":4.6,"publicationDate":"2024-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11026063/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140858506","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}
Pub Date : 2024-03-15eCollection Date: 2024-01-01DOI: 10.15698/mic2024.03.818
Fiorella Galello, Mariana Bermúdez-Moretti, María Clara Ortolá Martínez, Silvia Rossi, Paula Portela
The yeast Saccharomyces cerevisiae is widely used in food and non-food industries. During industrial fermentation yeast strains are exposed to fluctuations in oxygen concentration, osmotic pressure, pH, ethanol concentration, nutrient availability and temperature. Fermentation performance depends on the ability of the yeast strains to adapt to these changes. Suboptimal conditions trigger responses to the external stimuli to allow homeostasis to be maintained. Stress-specific signalling pathways are activated to coordinate changes in transcription, translation, protein function, and metabolic fluxes while a transient arrest of growth and cell cycle progression occur. cAMP-PKA, HOG-MAPK and CWI signalling pathways are turned on during stress response. Comprehension of the mechanisms involved in the responses and in the adaptation to these stresses during fermentation is key to improving this industrial process. The scope of this review is to outline the advancement of knowledge about the cAMP-PKA signalling and the crosstalk of this pathway with the CWI and HOG-MAPK cascades in response to the environmental challenges heat and hyperosmotic stress.
{"title":"The cAMP-PKA signalling crosstalks with CWI and HOG-MAPK pathways in yeast cell response to osmotic and thermal stress.","authors":"Fiorella Galello, Mariana Bermúdez-Moretti, María Clara Ortolá Martínez, Silvia Rossi, Paula Portela","doi":"10.15698/mic2024.03.818","DOIUrl":"10.15698/mic2024.03.818","url":null,"abstract":"<p><p>The yeast <i>Saccharomyces cerevisiae</i> is widely used in food and non-food industries. During industrial fermentation yeast strains are exposed to fluctuations in oxygen concentration, osmotic pressure, pH, ethanol concentration, nutrient availability and temperature. Fermentation performance depends on the ability of the yeast strains to adapt to these changes. Suboptimal conditions trigger responses to the external stimuli to allow homeostasis to be maintained. Stress-specific signalling pathways are activated to coordinate changes in transcription, translation, protein function, and metabolic fluxes while a transient arrest of growth and cell cycle progression occur. cAMP-PKA, HOG-MAPK and CWI signalling pathways are turned on during stress response. Comprehension of the mechanisms involved in the responses and in the adaptation to these stresses during fermentation is key to improving this industrial process. The scope of this review is to outline the advancement of knowledge about the cAMP-PKA signalling and the crosstalk of this pathway with the CWI and HOG-MAPK cascades in response to the environmental challenges heat and hyperosmotic stress.</p>","PeriodicalId":18397,"journal":{"name":"Microbial Cell","volume":"11 ","pages":"90-105"},"PeriodicalIF":4.6,"publicationDate":"2024-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10941952/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140143760","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}
Pub Date : 2024-03-14eCollection Date: 2024-01-01DOI: 10.15698/mic2024.03.817
Ivan Kushkevych, Kristýna Martínková, Lenka Mráková, Francesco Giudici, Simone Baldi, David Novak, Márió Gajdács, Monika Vítězová, Dani Dordevic, Amedeo Amedei, Simon K-M R Rittmann
Considerable evidence has accumulated regarding the molecular relationship between gut microbiota (GM) composition and the onset (clinical presentation and prognosis of ulcerative colitis (UC)). In addition, it is well documented that short-chain fatty acid (SCFA)-producing bacteria may play a fundamental role in maintaining an anti-inflammatory intestinal homeostasis, but sulfate- and sulfite reducing bacteria may be responsible for the production of toxic metabolites, such as hydrogen sulfide and acetate. Hence, the present study aimed to assess the GM composition - focusing on sulfate-reducing bacteria (SRB) - in patients with severe, severe-active and moderate UC. Each one of the six enrolled patients provided two stool samples in the following way: one sample was cultivated in a modified SRB-medium before 16S rRNA sequencing and the other was not cultivated. Comparative phylogenetic analysis was conducted on each sample. Percentage of detected gut microbial genera showed considerable variation based on the patients' disease severity and cultivation in the SRB medium. In detail, samples without cultivation from patients with moderate UC showed a high abundance of the genera Bacteroides, Bifidobacterium and Ruminococcus, but after SRB cultivation, the dominant genera were Bacteroides, Klebsiella and Bilophila. On the other hand, before SRB cultivation, the main represented genera in patients with severe UC were Escherichia-Shigella, Proteus, Methanothermobacter and Methanobacterium. However, after incubation in the SRB medium Bacteroides, Proteus, Alistipes and Lachnoclostridium were predominant. Information regarding GM compositional changes in UC patients may aid the development of novel therapeutic strategies (e.g., probiotic preparations containing specific bacterial strains) to counteract the mechanisms of virulence of harmful bacteria and the subsequent inflammatory response that is closely related to the pathogenesis of inflammatory bowel diseases.
{"title":"Comparison of microbial communities and the profile of sulfate-reducing bacteria in patients with ulcerative colitis and their association with bowel diseases: a pilot study.","authors":"Ivan Kushkevych, Kristýna Martínková, Lenka Mráková, Francesco Giudici, Simone Baldi, David Novak, Márió Gajdács, Monika Vítězová, Dani Dordevic, Amedeo Amedei, Simon K-M R Rittmann","doi":"10.15698/mic2024.03.817","DOIUrl":"10.15698/mic2024.03.817","url":null,"abstract":"<p><p>Considerable evidence has accumulated regarding the molecular relationship between gut microbiota (GM) composition and the onset (clinical presentation and prognosis of ulcerative colitis (UC)). In addition, it is well documented that short-chain fatty acid (SCFA)-producing bacteria may play a fundamental role in maintaining an anti-inflammatory intestinal homeostasis, but sulfate- and sulfite reducing bacteria may be responsible for the production of toxic metabolites, such as hydrogen sulfide and acetate. Hence, the present study aimed to assess the GM composition - focusing on sulfate-reducing bacteria (SRB) - in patients with severe, severe-active and moderate UC. Each one of the six enrolled patients provided two stool samples in the following way: one sample was cultivated in a modified SRB-medium before 16S rRNA sequencing and the other was not cultivated. Comparative phylogenetic analysis was conducted on each sample. Percentage of detected gut microbial genera showed considerable variation based on the patients' disease severity and cultivation in the SRB medium. In detail, samples without cultivation from patients with moderate UC showed a high abundance of the genera <i>Bacteroides</i>, <i>Bifidobacterium</i> and <i>Ruminococcus,</i> but after SRB cultivation, the dominant genera were <i>Bacteroides</i>, <i>Klebsiella</i> and <i>Bilophila</i>. On the other hand, before SRB cultivation, the main represented genera in patients with severe UC were <i>Escherichia-Shigella</i>, <i>Proteus</i>, <i>Methanothermobacter</i> and <i>Methanobacterium</i>. However, after incubation in the SRB medium <i>Bacteroides, Proteus, Alistipes</i> and <i>Lachnoclostridium</i> were predominant. Information regarding GM compositional changes in UC patients may aid the development of novel therapeutic strategies (e.g., probiotic preparations containing specific bacterial strains) to counteract the mechanisms of virulence of harmful bacteria and the subsequent inflammatory response that is closely related to the pathogenesis of inflammatory bowel diseases.</p>","PeriodicalId":18397,"journal":{"name":"Microbial Cell","volume":"11 ","pages":"79-89"},"PeriodicalIF":4.6,"publicationDate":"2024-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10939707/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140131865","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}
Pub Date : 2024-02-23eCollection Date: 2024-01-01DOI: 10.15698/mic2024.02.816
Jesús Gómez-Montalvo, Alvaro de Obeso Fernández Del Valle, Luis Fernando De la Cruz Gutiérrez, Jose Mario Gonzalez-Meljem, Christian Quintus Scheckhuber
Saccharomyces cerevisiae (baker's yeast) has yielded relevant insights into some of the basic mechanisms of organismal aging. Among these are genomic instability, oxidative stress, caloric restriction and mitochondrial dysfunction. Several genes are known to have an impact on the aging process, with corresponding mutants exhibiting short- or long-lived phenotypes. Research dedicated to unraveling the underlying cellular mechanisms can support the identification of conserved mechanisms of aging in other species. One of the hitherto less studied fields in yeast aging is how the organism regulates its gene expression at the transcriptional level. To our knowledge, we present the first investigation into alternative splicing, particularly intron retention, during replicative aging of S. cerevisiae. This was achieved by utilizing the IRFinder algorithm on a previously published RNA-seq data set by Janssens et al. (2015). In the present work, 44 differentially retained introns in 43 genes were identified during replicative aging. We found that genes with altered intron retention do not display significant changes in overall transcript levels. It was possible to functionally assign distinct groups of these genes to the cellular processes of mRNA processing and export (e.g., YRA1) in early and middle-aged yeast, and protein ubiquitination (e.g., UBC5) in older cells. In summary, our work uncovers a previously unexplored layer of the transcriptional program of yeast aging and, more generally, expands the knowledge on the occurrence of alternative splicing in baker's yeast.
{"title":"Replicative aging in yeast involves dynamic intron retention patterns associated with mRNA processing/export and protein ubiquitination.","authors":"Jesús Gómez-Montalvo, Alvaro de Obeso Fernández Del Valle, Luis Fernando De la Cruz Gutiérrez, Jose Mario Gonzalez-Meljem, Christian Quintus Scheckhuber","doi":"10.15698/mic2024.02.816","DOIUrl":"10.15698/mic2024.02.816","url":null,"abstract":"<p><p><i>Saccharomyces cerevisiae</i> (baker's yeast) has yielded relevant insights into some of the basic mechanisms of organismal aging. Among these are genomic instability, oxidative stress, caloric restriction and mitochondrial dysfunction. Several genes are known to have an impact on the aging process, with corresponding mutants exhibiting short- or long-lived phenotypes. Research dedicated to unraveling the underlying cellular mechanisms can support the identification of conserved mechanisms of aging in other species. One of the hitherto less studied fields in yeast aging is how the organism regulates its gene expression at the transcriptional level. To our knowledge, we present the first investigation into alternative splicing, particularly intron retention, during replicative aging of <i>S. cerevisiae</i>. This was achieved by utilizing the IRFinder algorithm on a previously published RNA-seq data set by Janssens <i>et al.</i> (2015). In the present work, 44 differentially retained introns in 43 genes were identified during replicative aging. We found that genes with altered intron retention do not display significant changes in overall transcript levels. It was possible to functionally assign distinct groups of these genes to the cellular processes of mRNA processing and export (e.g., <i>YRA1</i>) in early and middle-aged yeast, and protein ubiquitination (e.g., <i>UBC5</i>) in older cells. In summary, our work uncovers a previously unexplored layer of the transcriptional program of yeast aging and, more generally, expands the knowledge on the occurrence of alternative splicing in baker's yeast.</p>","PeriodicalId":18397,"journal":{"name":"Microbial Cell","volume":"11 ","pages":"69-78"},"PeriodicalIF":4.1,"publicationDate":"2024-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10897858/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139983301","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}
Pub Date : 2024-02-21eCollection Date: 2024-01-01DOI: 10.15698/mic2024.02.815
Thorsten Meyer, Oskar Knittelfelder, Martin Smolnig, Patrick Rockenfeller
Lipidomic analysis in diverse biological settings has become a frequent tool to increase our understanding of the processes of life. Cellular lipids play important roles not only as being the main components of cellular membranes, but also in the regulation of cell homeostasis as lipid signaling molecules. Yeast has been harnessed for biomedical research based on its good conservation of genetics and fundamental cell organisation principles and molecular pathways. Further application in so-called humanised yeast models have been developed which take advantage of yeast as providing the basics of a living cell with full control over heterologous expression. Here we present evidence that high-performance thin-layer chromatography (HPTLC) represents an effective alternative to replace cost intensive mass spectrometry-based lipidomic analyses. We provide statistical comparison of identical samples by both methods, which support the use of HPTLC for quantitative analysis of the main yeast lipid classes.
{"title":"Quantifying yeast lipidomics by high-performance thin-layer chromatography (HPTLC) and comparison to mass spectrometry-based shotgun lipidomics.","authors":"Thorsten Meyer, Oskar Knittelfelder, Martin Smolnig, Patrick Rockenfeller","doi":"10.15698/mic2024.02.815","DOIUrl":"10.15698/mic2024.02.815","url":null,"abstract":"<p><p>Lipidomic analysis in diverse biological settings has become a frequent tool to increase our understanding of the processes of life. Cellular lipids play important roles not only as being the main components of cellular membranes, but also in the regulation of cell homeostasis as lipid signaling molecules. Yeast has been harnessed for biomedical research based on its good conservation of genetics and fundamental cell organisation principles and molecular pathways. Further application in so-called humanised yeast models have been developed which take advantage of yeast as providing the basics of a living cell with full control over heterologous expression. Here we present evidence that high-performance thin-layer chromatography (HPTLC) represents an effective alternative to replace cost intensive mass spectrometry-based lipidomic analyses. We provide statistical comparison of identical samples by both methods, which support the use of HPTLC for quantitative analysis of the main yeast lipid classes.</p>","PeriodicalId":18397,"journal":{"name":"Microbial Cell","volume":"11 ","pages":"57-68"},"PeriodicalIF":4.6,"publicationDate":"2024-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10879857/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139931852","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}
Pub Date : 2024-02-20eCollection Date: 2024-01-01DOI: 10.15698/mic2024.02.814
Julia Graf, Leonard Fresenborg, Hans-Michael Seitz, Rafael Pernil, Enrico Schleiff
Metal homeostasis is central to all forms of life, as metals are essential micronutrients with toxic effects at elevated levels. Macromolecular machines facilitate metal uptake into the cells and their intracellular level is regulated by multiple means, which can involve RNA elements and proteinaceous components. While the general principles and components for uptake and cellular content regulation of, e.g., cobalt have been identified for proteobacteria, the corresponding mechanism in other Gram-negative bacteria such as cyanobacteria remain to be established. Based on their photosynthetic activity, cyanobacteria are known to exhibit a special metal demand in comparison to other bacteria. Here, the regulation by cobalt and cobalamin as well as their uptake is described for Anabaena sp. PCC 7120, a model filamentous heterocyst-forming cyanobacterium. Anabaena contains at least three cobalamin riboswitches in its genome, for one of which the functionality is confirmed here. Moreover, two outer membrane-localized cobalamin TonB-dependent transporters, namely BtuB1 and BtuB2, were identified. BtuB2 is important for fast uptake of cobalamin under conditions with low external cobalt, whereas BtuB1 appears to function in cobalamin uptake under conditions of sufficient cobalt supply. While the general function is comparable, the specific function of the two genes differs and mutants thereof show distinct phenotypes. The uptake of cobalamin depends further on the TonB and a BtuFCD machinery, as mutants of tonB3 and btuD show reduced cobalamin uptake rates. Thus, our results provide novel information on the uptake of cobalamin and the regulation of the cellular cobalt content in cyanobacteria.
{"title":"A cobalt concentration sensitive Btu-like system facilitates cobalamin uptake in <i>Anabaena</i> sp. PCC 7120.","authors":"Julia Graf, Leonard Fresenborg, Hans-Michael Seitz, Rafael Pernil, Enrico Schleiff","doi":"10.15698/mic2024.02.814","DOIUrl":"10.15698/mic2024.02.814","url":null,"abstract":"<p><p>Metal homeostasis is central to all forms of life, as metals are essential micronutrients with toxic effects at elevated levels. Macromolecular machines facilitate metal uptake into the cells and their intracellular level is regulated by multiple means, which can involve RNA elements and proteinaceous components. While the general principles and components for uptake and cellular content regulation of, e.g., cobalt have been identified for proteobacteria, the corresponding mechanism in other Gram-negative bacteria such as cyanobacteria remain to be established. Based on their photosynthetic activity, cyanobacteria are known to exhibit a special metal demand in comparison to other bacteria. Here, the regulation by cobalt and cobalamin as well as their uptake is described for <i>Anabaena</i> sp. PCC 7120, a model filamentous heterocyst-forming cyanobacterium. <i>Anabaena</i> contains at least three cobalamin riboswitches in its genome, for one of which the functionality is confirmed here. Moreover, two outer membrane-localized cobalamin TonB-dependent transporters, namely BtuB1 and BtuB2, were identified. BtuB2 is important for fast uptake of cobalamin under conditions with low external cobalt, whereas BtuB1 appears to function in cobalamin uptake under conditions of sufficient cobalt supply. While the general function is comparable, the specific function of the two genes differs and mutants thereof show distinct phenotypes. The uptake of cobalamin depends further on the TonB and a BtuFCD machinery, as mutants of <i>tonB3 and btuD</i> show reduced cobalamin uptake rates. Thus, our results provide novel information on the uptake of cobalamin and the regulation of the cellular cobalt content in cyanobacteria.</p>","PeriodicalId":18397,"journal":{"name":"Microbial Cell","volume":"11 ","pages":"41-56"},"PeriodicalIF":4.6,"publicationDate":"2024-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10878165/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139912948","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}
Pub Date : 2024-02-19eCollection Date: 2024-01-01DOI: 10.15698/mic2024.02.813
Shuo Shi, Yuwen Chu, Haiyan Liu, Lan Yu, Dejun Sun, Jialiang Yang, Geng Tian, Lei Ji, Cong Zhang, Xinxin Lu
Intratumoral microbiota can regulate the tumor immune microenvironment (TIME) and mediate tumor prognosis by promoting inflammatory response or inhibiting anti-tumor effects. Recent studies have elucidated the potential role of local tumor microbiota in the development and progression of lung adenocarcinoma (LUAD). However, whether intratumoral microbes are involved in the TIME that mediates the prognosis of LUAD remains unknown. Here, we obtained the matched tumor microbiome and host transcriptome and survival data of 478 patients with LUAD in The Cancer Genome Atlas (TCGA). Machine learning models based on immune cell marker genes can predict 1- to 5-year survival with relative accuracy. Patients were stratified into high- and low-survival-risk groups based on immune cell marker genes, with significant differences in intratumoral microbial communities. Specifically, patients in the high-risk group had significantly higher alpha diversity (p < 0.05) and were characterized by an enrichment of lung cancer-related genera such as Streptococcus. However, network analysis highlighted a more active pattern of dominant bacteria and immune cell crosstalk in TIME in the low-risk group compared to the high-risk group. Our study demonstrated that intratumoral microbiota-immune crosstalk was strongly associated with prognosis in LUAD patients, which would provide new targets for the development of precise therapeutic strategies.
瘤内微生物群可调节肿瘤免疫微环境(TIME),并通过促进炎症反应或抑制抗肿瘤作用来介导肿瘤预后。最近的研究阐明了局部肿瘤微生物群在肺腺癌(LUAD)发生和发展中的潜在作用。然而,瘤内微生物是否参与了介导 LUAD 预后的 TIME 仍是未知数。在此,我们从癌症基因组图谱(TCGA)中获得了478名LUAD患者的匹配肿瘤微生物组和宿主转录组及生存数据。基于免疫细胞标记基因的机器学习模型可以相对准确地预测1至5年的生存率。根据免疫细胞标记基因将患者分为高生存风险组和低生存风险组,肿瘤内微生物群落存在显著差异。具体来说,高风险组患者的α多样性明显更高(p < 0.05),其特点是富含链球菌等肺癌相关菌属。然而,网络分析显示,与高风险组相比,低风险组 TIME 中的优势菌和免疫细胞串扰模式更为活跃。我们的研究表明,瘤内微生物与免疫细胞间的串扰与肺癌患者的预后密切相关,这将为制定精确的治疗策略提供新的靶点。
{"title":"Predictable regulation of survival by intratumoral microbe-immune crosstalk in patients with lung adenocarcinoma.","authors":"Shuo Shi, Yuwen Chu, Haiyan Liu, Lan Yu, Dejun Sun, Jialiang Yang, Geng Tian, Lei Ji, Cong Zhang, Xinxin Lu","doi":"10.15698/mic2024.02.813","DOIUrl":"10.15698/mic2024.02.813","url":null,"abstract":"<p><p>Intratumoral microbiota can regulate the tumor immune microenvironment (TIME) and mediate tumor prognosis by promoting inflammatory response or inhibiting anti-tumor effects. Recent studies have elucidated the potential role of local tumor microbiota in the development and progression of lung adenocarcinoma (LUAD). However, whether intratumoral microbes are involved in the TIME that mediates the prognosis of LUAD remains unknown. Here, we obtained the matched tumor microbiome and host transcriptome and survival data of 478 patients with LUAD in The Cancer Genome Atlas (TCGA). Machine learning models based on immune cell marker genes can predict 1- to 5-year survival with relative accuracy. Patients were stratified into high- and low-survival-risk groups based on immune cell marker genes, with significant differences in intratumoral microbial communities. Specifically, patients in the high-risk group had significantly higher alpha diversity (p < 0.05) and were characterized by an enrichment of lung cancer-related genera such as <i>Streptococcus</i>. However, network analysis highlighted a more active pattern of dominant bacteria and immune cell crosstalk in TIME in the low-risk group compared to the high-risk group. Our study demonstrated that intratumoral microbiota-immune crosstalk was strongly associated with prognosis in LUAD patients, which would provide new targets for the development of precise therapeutic strategies.</p>","PeriodicalId":18397,"journal":{"name":"Microbial Cell","volume":"11 ","pages":"29-40"},"PeriodicalIF":4.6,"publicationDate":"2024-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10876218/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139905969","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}
Pub Date : 2024-01-09eCollection Date: 2024-01-01DOI: 10.15698/mic2024.01.812
Julia Graf, Martin Schöpperle, Rafael Pernil, Enrico Schleiff
Low availability of micronutrients such as iron has enforced the evolution of uptake systems in all kingdoms of life. In Gram-negative bacteria, outer membrane, periplasmatic and plasma membrane localized proteins facilitate the uptake of iron-loaded chelators, which are energized by TonB proteins. The specificity of different uptake systems likely depends either on the endogenously produced siderophore or on the bioavailability of iron-chelator complexes in the environment. Hence, an uptake system for schizokinen produced by the model cyanobacterium Anabaena sp. PCC 7120 exists, while bioinformatics analysis suggests the existence of additional systems, likely for uptake of xenosiderophores. Consistently, proteins encoded by alr2153 (hutA1) and alr3242 (hutA2) are assigned as outer membrane heme transporters. Indeed, Anabaena sp. PCC 7120 can utilize external heme as an iron source. The addition of heme resulted in an induction of genes involved in heme degradation and chlorophyll a synthesis and in an increase of the chlorophyll a content. Moreover, iron starvation induced the expression of hutA1, while the addition of heme led to its repression. Remarkably, the addition of a high concentration of heme but not iron starvation resulted in hutA2 induction. Plasmid insertion mutants of both genes exhibited a reduced capacity to recover from iron starvation by heme addition, which indicates a dependence of heme uptake on functional HutA1 and HutA2 proteins. The structural model generated by bioinformatics methods is further in agreement with a role in heme uptake. Thus, we provide evidence that Anabaena sp. PCC 7120 uses a heme uptake system in parallel to other iron acquisition systems.
{"title":"Two TonB-dependent outer membrane transporters involved in heme uptake in <i>Anabaena</i> sp. PCC 7120.","authors":"Julia Graf, Martin Schöpperle, Rafael Pernil, Enrico Schleiff","doi":"10.15698/mic2024.01.812","DOIUrl":"10.15698/mic2024.01.812","url":null,"abstract":"<p><p>Low availability of micronutrients such as iron has enforced the evolution of uptake systems in all kingdoms of life. In Gram-negative bacteria, outer membrane, periplasmatic and plasma membrane localized proteins facilitate the uptake of iron-loaded chelators, which are energized by TonB proteins. The specificity of different uptake systems likely depends either on the endogenously produced siderophore or on the bioavailability of iron-chelator complexes in the environment. Hence, an uptake system for schizokinen produced by the model cyanobacterium <i>Anabaena</i> sp. PCC 7120 exists, while bioinformatics analysis suggests the existence of additional systems, likely for uptake of xenosiderophores. Consistently, proteins encoded by <i>alr2153</i> (<i>hutA1</i>) and <i>alr3242</i> (<i>hutA2</i>) are assigned as outer membrane heme transporters. Indeed, <i>Anabaena</i> sp. PCC 7120 can utilize external heme as an iron source. The addition of heme resulted in an induction of genes involved in heme degradation and chlorophyll <i>a</i> synthesis and in an increase of the chlorophyll <i>a</i> content. Moreover, iron starvation induced the expression of <i>hutA1</i>, while the addition of heme led to its repression. Remarkably, the addition of a high concentration of heme but not iron starvation resulted in <i>hutA2</i> induction. Plasmid insertion mutants of both genes exhibited a reduced capacity to recover from iron starvation by heme addition, which indicates a dependence of heme uptake on functional HutA1 and HutA2 proteins. The structural model generated by bioinformatics methods is further in agreement with a role in heme uptake. Thus, we provide evidence that <i>Anabaena</i> sp. PCC 7120 uses a heme uptake system in parallel to other iron acquisition systems.</p>","PeriodicalId":18397,"journal":{"name":"Microbial Cell","volume":"11 ","pages":"16-28"},"PeriodicalIF":4.6,"publicationDate":"2024-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10792254/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139485630","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}
Pub Date : 2024-01-05eCollection Date: 2024-01-01DOI: 10.15698/mic2024.01.811
Yiannis Pyrris, Georgia F Papadaki, Emmanuel Mikros, George Diallinas
FurE is a H+ symporter specific for the cellular uptake of uric acid, allantoin, uracil, and toxic nucleobase analogues in the fungus Aspergillus nidulans. Being member of the NCS1 protein family, FurE is structurally related to the APC-superfamily of transporters. APC-type transporters are characterised by a 5+5 inverted repeat fold made of ten transmembrane segments (TMS1-10) and function through the rocking-bundle mechanism. Most APC-type transporters possess two extra C-terminal TMS segments (TMS11-12), the function of which remains elusive. Here we present a systematic mutational analysis of TMS11-12 of FurE and show that two specific aromatic residues in TMS12, Trp473 and Tyr484, are essential for ER-exit and trafficking to the plasma membrane (PM). Molecular modeling shows that Trp473 and Tyr484 might be essential through dynamic interactions with residues in TMS2 (Leu91), TMS3 (Phe111), TMS10 (Val404, Asp406) and other aromatic residues in TMS12. Genetic analysis confirms the essential role of Phe111, Asp406 and TMS12 aromatic residues in FurE ER-exit. We further show that co-expression of FurE-Y484F or FurE-W473A with wild-type FurE leads to a dominant negative phenotype, compatible with the concept that FurE molecules oligomerize or partition in specific microdomains to achieve concentrative ER-exit and traffic to the PM. Importantly, truncated FurE versions lacking TMS11-12 are unable to reproduce a negative effect on the trafficking of co-expressed wild-type FurE. Overall, we show that TMS11-12 acts as an intramolecular chaperone for proper FurE folding, which seems to provide a structural code for FurE partitioning in ER-exit sites.
{"title":"The last two transmembrane helices in the APC-type FurE transporter act as an intramolecular chaperone essential for concentrative ER-exit.","authors":"Yiannis Pyrris, Georgia F Papadaki, Emmanuel Mikros, George Diallinas","doi":"10.15698/mic2024.01.811","DOIUrl":"10.15698/mic2024.01.811","url":null,"abstract":"<p><p>FurE is a H<sup>+</sup> symporter specific for the cellular uptake of uric acid, allantoin, uracil, and toxic nucleobase analogues in the fungus <i>Aspergillus</i> nidulans. Being member of the NCS1 protein family, FurE is structurally related to the APC-superfamily of transporters. APC-type transporters are characterised by a 5+5 inverted repeat fold made of ten transmembrane segments (TMS1-10) and function through the rocking-bundle mechanism. Most APC-type transporters possess two extra C-terminal TMS segments (TMS11-12), the function of which remains elusive. Here we present a systematic mutational analysis of TMS11-12 of FurE and show that two specific aromatic residues in TMS12, Trp473 and Tyr484, are essential for ER-exit and trafficking to the plasma membrane (PM). Molecular modeling shows that Trp473 and Tyr484 might be essential through dynamic interactions with residues in TMS2 (Leu91), TMS3 (Phe111), TMS10 (Val404, Asp406) and other aromatic residues in TMS12. Genetic analysis confirms the essential role of Phe111, Asp406 and TMS12 aromatic residues in FurE ER-exit. We further show that co-expression of FurE-Y484F or FurE-W473A with wild-type FurE leads to a dominant negative phenotype, compatible with the concept that FurE molecules oligomerize or partition in specific microdomains to achieve concentrative ER-exit and traffic to the PM. Importantly, truncated FurE versions lacking TMS11-12 are unable to reproduce a negative effect on the trafficking of co-expressed wild-type FurE. Overall, we show that TMS11-12 acts as an intramolecular chaperone for proper FurE folding, which seems to provide a structural code for FurE partitioning in ER-exit sites.</p>","PeriodicalId":18397,"journal":{"name":"Microbial Cell","volume":"11 ","pages":"1-15"},"PeriodicalIF":4.6,"publicationDate":"2024-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10788122/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139472710","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}