Project description:To learn more about the global effects of phosphatidylcholine (PC)-deficiency in A. tumefaciens, we performed transcriptomic studies comparing the wild type and a PC-deficient DpmtA Dpcs mutant. We expanded the number of affected virulence genes and showed that the loss of PC was correlated with altered expression of multiple genes encoding membrane or membrane-associated proteins and proteins which fulfil their function at the membrane. The PC-deficient mutant analyzed in this study is further described in Sonja Klüsener, Stephanie Hacker, Yun-Long Tsai, Julia E. Bandow, Ronald Gust, Erh-Min Lai and Franz Narberhaus. 2010. Proteomic and transcriptomic characterization of a virulence-deficient phosphatidylcholine-negative Agrobacterium tumefaciens mutant.
Project description:We investigated the role of the LysR-type transcriptional regulator LsrB (Atu2186) from Agrobacterium tumefaciens C58 (alias A. fabrum C58), which regulates the expression of small-regulatory-RNA (sRNA) and several protein-coding genes, such as ampD (atu2113). In our current study we invesigated the role of LsrB in beta-lactam resistance. For this purpose, we constructed a vtlR/lsrB deletion mutant. Total RNA was isolated from this mutant. Sequencing was done on the Illumina NovaSeq 6000 platform. Wild-type samples that derived from the same experiment were published previously (Kraus et al., 2020) (GEO accession: GSE150941). In the vtlR/lsrB mutant around 800 protein coding genes and around 80 sRNA coding genes were downregulated or upregulated at least three-fold (p value ≤ 0.05). Overall, the results demonstrate the huge regulatory effect of LsrB on the transcriptome of A. tumefaciens. Kraus, A., Weskamp, M., Zierles, J., Balzer, M., Busch, R., Eisfeld, J., et al. (2020) Arginine-rich small proteins with a domain of unknown function DUF1127 play a role in phosphate and carbon metabolism of Agrobacterium tumefaciens. J Bacteriol 202: e00309-20.
Project description:To learn more about the global effects of phosphatidylcholine (PC)-deficiency in A. tumefaciens, we performed transcriptomic studies comparing the wild type and a PC-deficient DpmtA Dpcs mutant. We expanded the number of affected virulence genes and showed that the loss of PC was correlated with altered expression of multiple genes encoding membrane or membrane-associated proteins and proteins which fulfil their function at the membrane. The PC-deficient mutant analyzed in this study is further described in Sonja Klüsener, Stephanie Hacker, Yun-Long Tsai, Julia E. Bandow, Ronald Gust, Erh-Min Lai and Franz Narberhaus. 2010. Proteomic and transcriptomic characterization of a virulence-deficient phosphatidylcholine-negative Agrobacterium tumefaciens mutant. Total RNA was extracted from A. tumefaciens C58 wild-type and PC-deficient mutant cells grown under virulence-induced and non-induced conditions. The 4-plex NimbleGen Gene Expression Array for A. tumefaciens (Design ID 080630) represents 5344 genes and is based on NCBI reference sequences NC_003062, NC_003063, NC_003064 and NC_003065. Each gene is represented by 6 probes (45mer-60mer oligonucleotides), each in two copies on different locations on the array. Three biological replicates for each cultivation condition were analyzed for A. tumefaciens wild-type and PC-deficient mutant strains.
Project description:This research focuses on the design, manufacturing and validation of a new Agrobacterium tumefaciens C58 whole-genome tiling microarray platform for novel RNA transcript discovery. A whole-genome tiling microarray allows both annotated genes as well as previously unknown RNA transcripts to be detected and quantified at once. The Agrobacterium tumefaciens C58 genome is re-acquired with next-generation sequencing and then used to design the tilinlg microarray with the thermodynamic analysis program Picky. Validations are performed by subjecting Agrobacterium tumefaciens C58 under various growth conditions and then using the tling microarrays to verify expected gene expression patterns.