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  • 1. Cheng, J
    et al.
    Johansson, Magnus
    Nordlund, S
    Expression of P(II) and glutamine synthetase is regulated by P(II), the ntrBC products, and processing of the glnBA mRNA in Rhodospirillum rubrum1999In: Journal of Bacteriology, ISSN 0021-9193, E-ISSN 1098-5530, Vol. 181, no 20, p. 6530-6534Article in journal (Refereed)
    Abstract [en]

    We have studied the transcription of the glnB and glnA genes in Rhodospirillum rubrum with firefly luciferase as a reporter enzyme. Under NH(4)(+) and N(2) conditions, glnBA was cotranscribed from a weak and a strong promoter. In nitrogen-fixing cultures, activity of the latter was highly enhanced by NtrC, but transcription from both promoters occurred under both conditions. There is no promoter controlling transcription of glnA alone, supporting our proposal that the glnA mRNA is produced by processing.

  • 2. Johansson, Magnus
    et al.
    Nordlund, S
    Purification of P(II) and P(II)-UMP and in vitro studies of regulation of glutamine synthetase in Rhodospirillum rubrum1999In: Journal of Bacteriology, ISSN 0021-9193, E-ISSN 1098-5530, Vol. 181, no 20, p. 6524-6529Article in journal (Refereed)
    Abstract [en]

    The P(II) protein from Rhodospirillum rubrum was fused with a histidine tag, overexpressed in Escherichia coli, and purified by Ni(2+)-chelating chromatography. The uridylylated form of the P(II) protein could be generated in E. coli. The effects on the regulation of glutamine synthetase by P(II), P(II)-UMP, glutamine, and alpha-ketoglutarate were studied in extracts from R. rubrum grown under different conditions. P(II) and glutamine were shown to stimulate the ATP-dependent inactivation (adenylylation) of glutamine synthetase, which could be totally inhibited by alpha-ketoglutarate. Deadenylylation (activation) of glutamine synthetase required phosphate, but none of the effectors studied had any major effect, which is different from their role in the E. coli system. In addition, deadenylylation was found to be much slower than adenylylation under the conditions investigated.

  • 3. Johansson, Magnus
    et al.
    Nordlund, S
    Uridylylation of the P(II) protein in the photosynthetic bacterium Rhodospirillum rubrum1997In: Journal of Bacteriology, ISSN 0021-9193, E-ISSN 1098-5530, Vol. 179, no 13, p. 4190-4194Article in journal (Refereed)
    Abstract [en]

    The regulatory protein P(II) has been studied in great detail in enteric bacteria; however, its function in photosynthetic bacteria has not been clearly established. As a number of these bacteria have been shown to regulate nitrogenase activity by a metabolic control system, it is of special interest to establish the role of P(II) in these diazotrophs. In this study, we show that P(II) in Rhodospirillum rubrum is modified in response to the N status in the cell and that addition of ammonium or glutamine leads to demodification. We also provide evidence that P(II) is uridylylated. In addition, we show that not only these compounds but also NAD+ promotes demodification of P(II), which is of particular interest as this pyridine nucleotide has been shown to act as a switch-off effector of nitrogenase. Demodification of P(II) by ammonium or NAD+ did not occur in cultures treated with an inhibitor of glutamine synthetase (methionine sulfoximine), whereas treatment with the glutamate synthase inhibitor 6-diazo-5-oxo-norleucine led to total demodification of P(II) without any other addition. The results indicate that P(II) probably is not directly involved in darkness switch-off of nitrogenase but that a role in ammonium switch-off cannot be excluded.

  • 4.
    Lindroos, Hillevi L
    et al.
    Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Uppsala.
    Mira, Alex
    Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Uppsala; División de Microbiología, Universidad Miguel Hernández, Alicante, Spain.
    Repsilber, Dirk
    Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Uppsala; Institute for Medical Biometry and Statistics, University Lübeck, Lübeck, Germany .
    Vinnere, Olga
    Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Uppsala.
    Näslund, Kristina
    Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Uppsala.
    Dehio, Michaela
    Division of Molecular Microbiology, Biozentrum of the University of Basel, Basel, Switzerland .
    Dehio, Christoph
    Division of Molecular Microbiology, Biozentrum of the University of Basel, Basel, Switzerland .
    Andersson, Siv G E
    Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Uppsala.
    Characterization of the genome composition of Bartonella koehlerae by microarray comparative genomic hybridization profiling2005In: Journal of Bacteriology, ISSN 0021-9193, E-ISSN 1098-5530, Vol. 187, no 17, p. 6155-65Article in journal (Refereed)
    Abstract [en]

    Bartonella henselae is present in a wide range of wild and domestic feline hosts and causes cat-scratch disease and bacillary angiomatosis in humans. We have estimated here the gene content of Bartonella koehlerae, a novel species isolated from cats that was recently identified as an agent of human endocarditis. The investigation was accomplished by comparative genomic hybridization (CGH) to a microarray constructed from the sequenced 1.93-Mb genome of B. henselae. Control hybridizations of labeled DNA from the human pathogen Bartonella quintana with a reduced genome of 1.58 Mb were performed to evaluate the accuracy of the array for genes with known levels of sequence divergence. Genome size estimates of B. koehlerae by pulsed-field gel electrophoresis matched that calculated by the CGH, indicating a genome of 1.7 to 1.8 Mb with few unique genes. As in B. quintana, sequences in the prophage and the genomic islands were reported absent in B. koehlerae. In addition, sequence variability was recorded in the chromosome II-like region, where B. koehlerae showed an intermediate retention pattern of both coding and noncoding sequences. Although most of the genes missing in B. koehlerae are also absent from B. quintana, its phylogenetic placement near B. henselae suggests independent deletion events, indicating that host specificity is not solely attributed to genes in the genomic islands. Rather, the results underscore the instability of the genomic islands even within bacterial populations adapted to the same host-vector system, as in the case of B. henselae and B. koehlerae.

  • 5.
    Lindroos, Hillevi
    et al.
    Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Uppsala .
    Vinnere, Olga
    Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Uppsala.
    Mira, Alex
    Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Uppsala.
    Repsilber, Dirk
    Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Uppsala.
    Näslund, Kristina
    Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Uppsala.
    Andersson, Siv G E
    Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Uppsala.
    Genome rearrangements, deletions, and amplifications in the natural population of Bartonella henselae2006In: Journal of Bacteriology, ISSN 0021-9193, E-ISSN 1098-5530, Vol. 188, no 21, p. 7426-39Article in journal (Refereed)
    Abstract [en]

    Cats are the natural host for Bartonella henselae, an opportunistic human pathogen and the agent of cat scratch disease. Here, we have analyzed the natural variation in gene content and genome structure of 38 Bartonella henselae strains isolated from cats and humans by comparative genome hybridizations to microarrays and probe hybridizations to pulsed-field gel electrophoresis (PFGE) blots. The variation in gene content was modest and confined to the prophage and the genomic islands, whereas the PFGE analyses indicated extensive rearrangements across the terminus of replication with breakpoints in areas of the genomic islands. We observed no difference in gene content or structure between feline and human strains. Rather, the results suggest multiple sources of human infection from feline B. henselae strains of diverse genotypes. Additionally, the microarray hybridizations revealed DNA amplification in some strains in the so-called chromosome II-like region. The amplified segments were centered at a position corresponding to a putative phage replication initiation site and increased in size with the duration of cultivation. We hypothesize that the variable gene pool in the B. henselae population plays an important role in the establishment of long-term persistent infection in the natural host by promoting antigenic variation and escape from the host immune response.

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