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@PHDTHESIS{Pahlke:154827,
      author       = {Pahlke, Jennifer},
      title        = {{T}he 6 {RNA} of $\textit{{C}orynebacterium glutamicum}$},
      volume       = {76},
      school       = {Heinrich-Heine-Universität Düsseldorf},
      type         = {Dr.},
      address      = {Jülich},
      publisher    = {Forschungszentrum Jülich GmbH Zentralbibliothek, Verlag},
      reportid     = {FZJ-2014-04095},
      isbn         = {978-3-95806-003-6},
      series       = {Schriften des Forschungszentrums Jülich. Reihe Gesundheit
                      / Health},
      pages        = {II, 144 S.},
      year         = {2014},
      note         = {Biotechnologie 1, nicht OA!!!; Heinrich-Heine-Universität
                      Düsseldorf, Diss., 2014},
      abstract     = {The 6C RNA family is a class of small RNAs (sRNAs) with as
                      yet unknown function present and conserved within the phylum
                      Actinobacteria, including the Gram-positive soil bacterium
                      $\textit{Corynebacterium glutamicum}$. In this work studies
                      were conducted to characterize properties of the 6C RNA from
                      $\textit{C. glutamicum}$ ATCC 13032 and to shed light on
                      possible physiological roles in this species. The following
                      results were obtained: (i) The 6C RNA was found to be
                      present in $\textit{C. glutamicum}$ throughout growth in
                      CGXII medium with glucose and exhibited RNA level changes
                      not greater than two-fold. RNA stability tests using
                      rifampicin revealed a 6C RNA half-life of >120 min, showing
                      that it is a very stable sRNA. Quantitation of the 6C RNA
                      transcripts yielded about 325 and about 260 molecules per
                      cell in the exponential and in the stationary growth phase,
                      respectively, which is much more compared to numbers of
                      individual mRNAs. (ii) 6C RNA pull-down experiments using
                      crude protein extracts of $\textit{C. glutamicum}$ revealed
                      3-phosphoglycerate kinase (PGK) as a candidate protein
                      interaction partner. However, an interaction of 6C RNA with
                      purified PGK could not be confirmed yet by electrophoretic
                      mobility shift assays, possibly indicating that a further
                      factor is required. (iii) The 6C RNA is not essential for
                      $\textit{C. glutamicum}$ and the growth of a 6C RNA deletion
                      mutant was very similar to the wild type under various
                      cultivation conditions tested, except when treated with the
                      SOS response-inducing antibiotic mitomycin C (MMC), which
                      caused a growth defect of the Δ6C RNA mutant. Reporter
                      fusion constructs with promoters of the LexA-regulated genes
                      $\textit{recA, recN, cglIM}$ and $\textit{dnaE2}$ indicated
                      that the SOS response level in the Δ6C RNA mutant is lower
                      in the stationary growth phase compared to the wild type.
                      This leads to the suggestion that the 6C RNA might be
                      involved in the SOS response or plays a more general role in
                      the stress defense of $\textit{C. glutamicum}$.
                      Complementation studies of the Δ6C RNA mutant by
                      plasmid-born expression of the 6C RNA from its native
                      promoter revealed an unexpected and exceptional growth
                      phenotype in the presence of MMC. Quantitation of 6C RNA
                      revealed an about five-fold increased level in the
                      complemented strain. (iv) The increased 6C RNA level
                      resulted in an elongated cell shape under standard
                      conditions and a dramatically branched cell shape in the
                      presence of MMC, indicating that the 6C RNA might be
                      involved in the regulation of cell division in $\textit{C.
                      glutamicum}$. (v) 6C RNA deletion as well as 6C RNA
                      overexpression led to changes in the global mRNA expression
                      profile. Notably, the WhiB-like transcriptional regulator
                      WhcD had a decreased mRNA level in the 6C RNA overproducer.
                      WhcD is homologous to the septation and cell
                      division-related regulator WhmD in $\textit{Mycobacterium
                      smegmatis}$. A Δ$\textit{whcD}$ mutant showed a reduced
                      growth and a strongly altered cell morphology resembling
                      that of the 6C RNA overproducer in the presence of MMC.
                      Thus, the homologous proteins indeed seem to be functionally
                      equivalent in both actinobacteria. These results suggest
                      that the 6C RNA may have an impact on the expression of
                      $\textit{whcD}$ or its mRNA stability, thereby playing a
                      role in some aspect of septation and cell division in
                      $\textit{C. glutamicum}$.},
      keywords     = {Dissertation (GND)},
      cin          = {IBG-1},
      cid          = {I:(DE-Juel1)IBG-1-20101118},
      pnm          = {899 - ohne Topic (POF2-899)},
      pid          = {G:(DE-HGF)POF2-899},
      typ          = {PUB:(DE-HGF)11},
      url          = {https://juser.fz-juelich.de/record/154827},
}