Amelioration of plasmid carriage
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Project description:pQBR103 is a naturally occurring plasmid known to alter the behaviour of its host, Pseudomonas fluorescens. This plasmid encodes a predicted translational regulator, rsmQ which is a homologue of known global regulators within Pseudomonas. Within this study we compared Pseudomonas fluorescens SBW25 plasmid free cells with cells carrying either WT pQBR103 or pQBR103 lacking rsmQ. Using comparative quantitative proteomics and RNAseq we showed that the loss of rsmQ leads to widespread proteomic changes in the absence of changes in mRNA abundance, suggesting RsmQ is a translational regulator of its host chromosome.
2023-01-13 | PXD033640 | Pride
Project description:Plasmid carriage requires appropriate expression of the genes on the plasmid or host chromosome through cooperative transcriptional regulation. To clarify the impact of plasmid carriage on the host chromosome, we compared the chromosomal RNA maps of plasmid-free and plasmid-containing host strains using the incompatibility group P-7 archetype plasmid pCAR1, which is involved in carbazole degradation, and three distinct Pseudomonas strains. The possession of pCAR1 altered gene expression related to the iron acquisition systems in each host. Expression of the major siderophore pyoverdine was greater in plasmid-containing P. putida KT2440 and P. aeruginosa PAO1 than in the plasmid-free host strains, in part due to the expression of carbazole-degradative genes on pCAR1. The mexEFoprN operon encoding an efflux pump of the resistance-nodulation-cell division (RND) family was specifically upregulated by the carriage of pCAR1 in P. putida KT2440, whereas the expression of orthologous genes in the other species remained unaltered. Induction of the mexEFoprN genes increased the resistance of pCAR1-containing KT2440 to chloramphenicol compared to pCAR1-free KT2440. Our findings indicate that the possession of pCAR1 altered the growth rate of the host via the expression of genes on pCAR1 and the host chromosomes.
2009-12-25 | GSE15217 | GEO
Project description:Plasmids can affect expression of genes on the host chromosome. This experiment was carried out to determine the effect of the presence of the plasmid pQBR103 (with and without the rsmQ gene on it) on global gene expression in Pseudomonas fluorescens SBW25.
2022-12-15 | E-MTAB-11868 | biostudies-arrayexpress
Project description:Impacts of plasmid carriage on its host cell were comprehensively analyzed using conjugative plasmid pCAR1 in the three different kinds of hosts, Pseudomonas putida KT2440, P. aeruginosa PAO1, and P. fluorescens Pf0-1. Various analyses of the host phenotype showed that pCAR1 carriage reduced host fitness, swimming motility, and resistances to osmotic- or pH-stress, and brought about the alterations of primary metabolic capacities in the TCA cycle or those several steps away from the TCA cycle in the host cells. Growth phase-dependent transcriptome analyses were performed with the classification of the annotated genes based on their identities among the three hosts and their putative functions. pCAR1 carriage affected host transcriptome more greatly at the transition and stationary phases in each host. The transcriptome responses were more similar between KT2440 and PAO1 than between other host combinations, and many genes, such as for ribosomal proteins, F-type ATPase, and RNAP core, in both strains were commonly not suppressed in their stationary phases. These responses may have resulted in the reduction of host fitness, motility, and stress resistances. Host-specific responses to plasmid carriage were transcriptional changes of genes on putative prophage or foreign DNA regions. The extent of the impacts in host phenotypes and transcriptomes was similarly the largest in KT2440 and the lowest in Pf0-1. The host alterations controlled by pCAR1 carriage are important for understanding the fate of the plasmid and its host, plasmid maintenance, expression of plasmid genes, the host cell physiology, and host survivability in the environment.
2012-12-25 | GSE39636 | GEO
Project description:Plasmids are one of the important mobile genetic elements in bacterial evolution. In this study, to evaluate the generality of the impact of plasmid carriage on host cell between different plasmids, we compared the response of Pseudomonas putida KT2440 to harboring three natural plasmids; RP4 (IncP-1, multidrug resistance, 60,099-bp), pCAR1 (IncP-7, carbazole-degradative, 200,231-bp) and NAH7 (IncP-9, naphthalene-degradative, 82,232-bp). We prepared two sets of plasmid-harboring strains from independent conjugation events to elucidate the reproducibility of the impact of the plasmid carriage. As results, the fitness was reduced by the carriage of RP4 and pCAR1 in liquid medium, while it was unaffected or even improved for NAH7-harboring strains. RP4-harboring KT2440 formed smaller colonies than the plasmid-free strain on solid medium (1.6% agar). The host cells were elongated by the carriage of the all plasmids, respectively. Copy number determination by quantitative PCR showed that the amount of each plasmid DNA in the host cell did not differed drastically. Whole genome resequencing showed that 13 SNPs (RP4), 24 SNPs (pCAR1) and 5 SNPs (NAH7) were the total differences between the two substrains for each plasmid-harboring strains. Transcriptome analyses showed that the impact of plasmid carriage was constantly larger in RP4-harboring strain than the other two plasmid-harboring strains. Genes involved in metal acquisition and metabolism were commonly affected by the carriage of the three plasmid. Indeed, plasmid-harboring strains showed greater growth inhibition than plasmid-free strains under iron-limiting condition. This feature could become future target to control plasmid spreading.
2018-02-17 | GSE110733 | GEO
Project description:Impacts of plasmid carriage on its host cell were comprehensively analyzed using conjugative plasmid pCAR1 in the three different kinds of hosts, Pseudomonas putida KT2440, P. aeruginosa PAO1, and P. fluorescens Pf0-1. Various analyses of the host phenotype showed that pCAR1 carriage reduced host fitness, swimming motility, and resistances to osmotic- or pH-stress, and brought about the alterations of primary metabolic capacities in the TCA cycle or those several steps away from the TCA cycle in the host cells. Growth phase-dependent transcriptome analyses were performed with the classification of the annotated genes based on their identities among the three hosts and their putative functions. pCAR1 carriage affected host transcriptome more greatly at the transition and stationary phases in each host. The transcriptome responses were more similar between KT2440 and PAO1 than between other host combinations, and many genes, such as for ribosomal proteins, F-type ATPase, and RNAP core, in both strains were commonly not suppressed in their stationary phases. These responses may have resulted in the reduction of host fitness, motility, and stress resistances. Host-specific responses to plasmid carriage were transcriptional changes of genes on putative prophage or foreign DNA regions. The extent of the impacts in host phenotypes and transcriptomes was similarly the largest in KT2440 and the lowest in Pf0-1. The host alterations controlled by pCAR1 carriage are important for understanding the fate of the plasmid and its host, plasmid maintenance, expression of plasmid genes, the host cell physiology, and host survivability in the environment. The growth-dependent change of chromosomal RNA maps from exponential to early stationary phases of pCAR1-harboring KT2440 and non-pCAR1 harboring KT2440 were compared. The growth-dependent change of chromosomal RNA maps from exponential to early stationary phases of pCAR1-harboring PAO1 and non-pCAR1 harboring PAO1 were compared. The growth-dependent change of chromosomal RNA maps from exponential to early stationary phases of pCAR1-harboring Pf0-1 and non-pCAR1 harboring Pf0-1 were compared. The growth-dependent change of RNA maps from exponential to early stationary phases of plasmid pCAR1possessed by 3 kinds of hosts were compared.
2012-12-25 | E-GEOD-39636 | biostudies-arrayexpress
Project description:Aim: to identify the transcriptional responses to acquisition of conjugative megaplasmids by Pseudomonas fluorescens, and how these effects are impacted by known compensatory mutations. Full abstract TBC.
2021-04-17 | GSE151570 | GEO
Project description:To elucidate the impact of the carriage of carbazole-degradative plasmid pCAR1 on biofilm formation by host bacteria, we compared biofilm morphology of pCAR1-free and pCAR1-carrying three Pseudomonas hosts: P. putida KT2440, P. aeruginosa PAO1 and P. fluorescens Pf0-1 using confocal laser scanning microscopy. Although pCAR1 carriage had no significant influence on biofilm formed by PAO1 and Pf0-1, pCAR1-carrying KT2440 became filamentous and formed flat biofilm, whereas pCAR1-free KT2440 formed mushroom-like biofilm. pCAR1 carries three genes encoding nucleoid-associated proteins (NAPs); Pmr, Pnd, and Phu. Because intensive filamentous morphology was observed in two double mutants (DpmrDpnd and DpmrDphu), these NAPs cooperatively suppress cell filamentation and flat biofilm formation in KT2440(pCAR1) cells. Based on the results of transcriptome analyses of these double mutants, we could find 32 candidate genes which may be involved in the filamentation of KT2440(pCAR1). Overproduction in KT2440 and KT2440(pCAR1) of three of candidate genes (PP_2193, PP_0308, or PP_0309) induced temporal filamentation of the cells, suggesting that pCAR1 induced the abnormal filamentous morphology of KT2440 cells by overexpression of plural genes including PP_2193, PP_0308, and PP_0309. In addition, pCAR1-borne NAPs suppress the morphological changes probably by decreasing the transcriptome change by pCAR1 carriage. The pCAR1 carriage impact of chromosomal RNA maps in biofilm formation.
2014-05-08 | E-GEOD-56856 | biostudies-arrayexpress
Project description:To elucidate the impact of the carriage of carbazole-degradative plasmid pCAR1 on biofilm formation by host bacteria, we compared biofilm morphology of pCAR1-free and pCAR1-carrying three Pseudomonas hosts: P. putida KT2440, P. aeruginosa PAO1 and P. fluorescens Pf0-1 using confocal laser scanning microscopy. Although pCAR1 carriage had no significant influence on biofilm formed by PAO1 and Pf0-1, pCAR1-carrying KT2440 became filamentous and formed flat biofilm, whereas pCAR1-free KT2440 formed mushroom-like biofilm. pCAR1 carries three genes encoding nucleoid-associated proteins (NAPs); Pmr, Pnd, and Phu. Because intensive filamentous morphology was observed in two double mutants (DpmrDpnd and DpmrDphu), these NAPs cooperatively suppress cell filamentation and flat biofilm formation in KT2440(pCAR1) cells. Based on the results of transcriptome analyses of these double mutants, we could find 32 candidate genes which may be involved in the filamentation of KT2440(pCAR1). Overproduction in KT2440 and KT2440(pCAR1) of three of candidate genes (PP_2193, PP_0308, or PP_0309) induced temporal filamentation of the cells, suggesting that pCAR1 induced the abnormal filamentous morphology of KT2440 cells by overexpression of plural genes including PP_2193, PP_0308, and PP_0309. In addition, pCAR1-borne NAPs suppress the morphological changes probably by decreasing the transcriptome change by pCAR1 carriage.
2014-05-08 | GSE56856 | GEO
Project description:Pseudomonas species are ubiquitous in plant-associated environments and produce an array of volatiles, enzymes and antimicrobials. The biosynthesis of many metabolites is regulated by the GacS/GacA two-component regulatory system. Transcriptome analysis of Pseudomonas fluorescens SBW25 revealed that 702 genes were differentially regulated (fold change>4, P<0.0001) in a gacS::Tn5 mutant, with 300 and 402 genes up- and down-regulated, respectively. Genes that were significantly down-regulated are involved in viscosin biosynthesis (viscABC), protease production (aprA), motility, biofilm formation, and secretory systems. Genes that were significantly up-regulated are involved in siderophore biosynthesis and oxidative stress. In contrast to previous studies with gac-mutants of other Pseudomonas species/strains, the gacS mutant of SBW25 inhibited growth of oomycete, fungal and bacterial pathogens significantly more than parental strain SBW25. A potential candidate for this enhanced antimicrobial activity was a large nonribosomal peptide synthetase (NRPS) gene cluster predicted to encode for an 8-amino-acid ornicorrugatin-like peptide. Site-directed mutagenesis of an NRPS gene in this cluster, however, did not lead to a reduction in the antimicrobial activity of the gacS mutant. Collectively these results indicate that a mutation in the GacS/GacA regulatory system causes major transcriptional changes in P. fluorescens SBW25 and significantly enhances its antimicrobial activities by yet unknown mechanisms. This expression study used total RNA recovered from four separate wild-type cultures of Pseudomonas fluorescens SBW25 and four separate cultures of the gacS mutant. Expression design was based on the updated genome sequence of Pseudomonas fluorescens SBW25, NC_012660.1 and associated plasmid pQBR0476 with nineteen 60-mer probe per gene. Each probe is replicated 3 times. The design includes random GC and other control probes.
2013-06-17 | E-GEOD-43443 | biostudies-arrayexpress