Project description:This project is about the proteomic and phosphoproteomic analyses of phage JSS1 or JSS1Δ004 infected Salmonella strains. AD032TQ is proteomes of phage JSS1 or JSS1Δ004 infected S. enterica serovar Cerro 87. AD032TPST is phosphoproteome of phage JSS1 or JSS1Δ004 infected S. enterica serovar Cerro 87. AE065TQ is proteomes of phage JSS1 or JSS1Δ004 infected CX1, a S. enterica serovar Cerro 87 dndFGH deficient strain, harboring a plasmid expressing DndFGH from Escherichia coli B7A. AE065TPST is phosphoproteome of phage JSS1 or JSS1Δ004 infected CX1 harboring a plasmid expressing DndFGH from E. coli B7A.
Project description:This project is about the proteomic and phosphoproteomic analyses of phage JSS1 or JSS1Δ004 infected Salmonella strains. AD032TQ is proteomes of phage JSS1 or JSS1Δ004 infected S. enterica serovar Cerro 87. AD032TPST is phosphoproteome of phage JSS1 or JSS1Δ004 infected S. enterica serovar Cerro 87. AE065TQ is proteomes of phage JSS1 or JSS1Δ004 infected CX1, a S. enterica serovar Cerro 87 dndFGH deficient strain, harboring a plasmid expressing DndFGH from Escherichia coli B7A. AE065TPST is phosphoproteome of phage JSS1 or JSS1Δ004 infected CX1 harboring a plasmid expressing DndFGH from E. coli B7A.
Project description:This project is about the proteomic and phosphoproteomic analyses of phage JSS1 or JSS1Δ004 infected Salmonella strains. AD032TQ is proteomes of phage JSS1 or JSS1Δ004 infected S. enterica serovar Cerro 87. AD032TPST is phosphoproteome of phage JSS1 or JSS1Δ004 infected S. enterica serovar Cerro 87. AE065TQ is proteomes of phage JSS1 or JSS1Δ004 infected CX1, a S. enterica serovar Cerro 87 dndFGH deficient strain, harboring a plasmid expressing DndFGH from Escherichia coli B7A. AE065TPST is phosphoproteome of phage JSS1 or JSS1Δ004 infected CX1 harboring a plasmid expressing DndFGH from E. coli B7A.
Project description:This project is about the proteomic and phosphoproteomic analyses of phage JSS1 or JSS1Δ004 infected Salmonella strains. AD032TQ is proteomes of phage JSS1 or JSS1Δ004 infected S. enterica serovar Cerro 87. AD032TPST is phosphoproteome of phage JSS1 or JSS1Δ004 infected S. enterica serovar Cerro 87. AE065TQ is proteomes of phage JSS1 or JSS1Δ004 infected CX1, a S. enterica serovar Cerro 87 dndFGH deficient strain, harboring a plasmid expressing DndFGH from Escherichia coli B7A. AE065TPST is phosphoproteome of phage JSS1 or JSS1Δ004 infected CX1 harboring a plasmid expressing DndFGH from E. coli B7A.
Project description:Compared with HKE9, HKE9-M-rpoS (rpoS knockout in hypervirulent Klebsiella pneumoniae strain HKE9) had 624 genes expressed significantly different, among which 148 genes were significantly up-regulated and 476 genes were significantly down-regulated, 586 genes on chromosomes, 10 genes on circular plasmid and 28 genes on linear plasmid; HKE9-C-M-rpoS had 746 genes expressed significantly different, among which 286 genes were significantly up-regulated and 460 genes were significantly down-regulated, 727 genes on chromosomes, 11 genes on circular plasmid and 8 genes on linear plasmid. Compared with HKE9-M-rpoS, HKE9-C-M-rpoS (rpoS complement in strain HKE9-M-rpoS) had 509 genes expressed significantly different, among which 324 genes were significantly up-regulated and 185 genes were significantly down-regulated, 458 genes on chromosomes, 3 genes on circular plasmid and 48 genes on linear plasmid.
Project description:Low transfection efficiency poses a significant challenge to experimental success, making the improvement of gene-carrying plasmid delivery and expression essential across various research fields. The stability of transfected vector DNA or mRNA significantly impacts transgene expression, as the innate immune system may recognize plasmid DNA as foreign and initiate a degradation response. Currently, it is unclear whether RNA transcribed from plasmids can activate RNA sensors, affecting transfection efficiency. This study employed RNA sequencing to analyze cellular responses to various circular and linear DNAs across five cell lines—HEK293T, HCT116, HeLa, L02, and NCM460—revealing that the innate immune response is a primary contributor to low transfection efficiency. Additionally, we used ChIP-seq to detect H3K27ac histone marks to explore whether the immune response triggered by plasmid transfection is regulated by epigenetic mechanisms. Finally, we performed ChIP assays for IRF7 and p-IRF3 in HeLa cells transfected with the linear pcDNA3.1-neo plasmid to demonstrate that these two transcription factors can indeed bind to the RTV genes.
Project description:Lyme disease spirochetes must induce RpoS-dependent genes during tick feeding to prepare for host infection. Previous work in our lab identified bbd18 as a negative regulator of RpoS, but inactivation of bbd18 in wild-type spirochetes was never achieved. In the current study we generated an inducible bbd18 gene at the endogenous plasmid locus and demonstrated the essential nature of BBD18 for viability of wild-type spirochetes in vitro and at a unique point in vivo. Transcriptomic analyses demonstrated global induction of RpoS and RpoS-dependent genes following BBD18 depletion, culminating in spirochete lysis. Plasmid prophage genes were also induced and phage particles were detected in lysed culture supernatants, suggesting that RpoS regulates phage lysis-lysogeny decisions. The absolute requirement for BBD18 persisted following displacement of the entire set of cp32 plasmid prophages but could be circumvented by deletion of rpoS. This is the first report of a mechanistic link between endogenous transducing prophages and the RpoS-dependent adaptive response of the Lyme disease spirochete.
Project description:Active segregation of DNA in bacteria is catalyzed by cytomotive structures. Mediators of subcellular plasmid positioning are Walker-type ATPases (ParA), actin-like proteins, or tubulin homologs. These motor proteins are coupled to the DNA via adaptor proteins that recognize specific DNA motifs. Here, we describe that a temperate phage, CGP3, integrated into the genome of Corynebacterium glutamicum ATCC 13032 encodes an actin-like protein, AlpC. Biochemical characterization confirms that AlpC is a bona fide actin-like protein and cell biological analysis shows that AlpC forms dynamic filamentous structures upon phage induction. The co-transcribed AlpA protein binds to a specific region of the phage DNA, possibly functioning as an adaptor protein that connects circular phage DNA to the tips of the AlpC filaments. The AlpC filaments transport phage DNA to the cell membrane of the host cell. Furthermore, both AlpA and AlpC are required for efficient phage replication. This is remarkably similar to actin-assisted membrane localization of eukaryotic viruses that use the actin cytoskeleton to concentrate virus particles at the egress sites.
Project description:Active segregation of DNA in bacteria is catalyzed by cytomotive structures. Mediators of subcellular plasmid positioning are Walker-type ATPases (ParA), actin-like proteins, or tubulin homologs. These motor proteins are coupled to the DNA via adaptor proteins that recognize specific DNA motifs. Here, we describe that a temperate phage, CGP3, integrated into the genome of Corynebacterium glutamicum ATCC 13032 encodes an actin-like protein, AlpC. Biochemical characterization confirms that AlpC is a bona fide actin-like protein and cell biological analysis shows that AlpC forms dynamic filamentous structures upon phage induction. The co-transcribed AlpA protein binds to a specific region of the phage DNA, possibly functioning as an adaptor protein that connects circular phage DNA to the tips of the AlpC filaments. The AlpC filaments transport phage DNA to the cell membrane of the host cell. Furthermore, both AlpA and AlpC are required for efficient phage replication. This is remarkably similar to actin-assisted membrane localization of eukaryotic viruses that use the actin cytoskeleton to concentrate virus particles at the egress sites.