Recent Submissions

  • YY1-Binding Sites Provide Central Switch Functions in the PARP-1 Gene Expression Network.

    Doetsch, Martina; Gluch, Angela; Poznanović, Goran; Bode, Juergen; Vidaković, Melita; Helmholtz Centre for Infection Research/Epigenetic Regulation, Braunschweig, Germany. (2012)
    Evidence is presented for the involvement of the interplay between transcription factor Yin Yang 1 (YY1) and poly(ADP-ribose) polymerase-1 (PARP-1) in the regulation of mouse PARP-1 gene (muPARP-1) promoter activity. We identified potential YY1 binding motifs (BM) at seven positions in the muPARP-1 core-promoter (-574/+200). Binding of YY1 was observed by the electrophoretic supershift assay using anti-YY1 antibody and linearized or supercoiled forms of plasmids bearing the core promoter, as well as with 30 bp oligonucleotide probes containing the individual YY1 binding motifs and four muPARP-1 promoter fragments. We detected YY1 binding to BM1 (-587/-558), BM4 (-348/-319) and a very prominent association with BM7 (+86/+115). Inspection of BM7 reveals overlap of the muPARP-1 translation start site with the Kozak sequence and YY1 and PARP-1 recognition sites. Site-directed mutagenesis of the YY1 and PARP-1 core motifs eliminated protein binding and showed that YY1 mediates PARP-1 binding next to the Kozak sequence. Transfection experiments with a reporter gene under the control of the muPARP-1 promoter revealed that YY1 binding to BM1 and BM4 independently repressed the promoter. Mutations at these sites prevented YY1 binding, allowing for increased reporter gene activity. In PARP-1 knockout cells subjected to PARP-1 overexpression, effects similar to YY1 became apparent; over expression of YY1 and PARP-1 revealed their synergistic action. Together with our previous findings these results expand the PARP-1 autoregulatory loop principle by YY1 actions, implying rigid limitation of muPARP-1 expression. The joint actions of PARP-1 and YY1 emerge as important contributions to cell homeostasis.
  • Novel tag-and-exchange (RMCE) strategies generate master cell clones with predictable and stable transgene expression properties.

    Qiao, Junhua; Oumard, André; Wegloehner, Wolfgang; Bode, Juergen; Department of Molecular Biotechnology/Epigenetic Regulation, Helmholtz Centre for Infection Research, Braunschweig, Germany. (2009-07-24)
    Site-specific recombinases have revolutionized the systematic generation of transgenic cell lines and embryonic stem cells/animals and will ultimately also reveal their potential in the genetic modification of induced pluripotent stem cells. Introduced in 1994, our Flp recombinase-mediated cassette exchange strategy permits the exchange of a target cassette for a cassette with the gene of interest, introduced as a part of an exchange vector. The process is "clean" in the sense that it does not co-introduce prokaryotic vector parts; neither does it leave behind a selection marker. Stringent selection principles provide master cell lines permitting subsequent recombinase-mediated cassette exchange cycles in the absence of a drug selection and with a considerable efficiency (approximately 10%). Exemplified by Chinese hamster ovary cells, the strategy proves to be successful even for cell lines with an unstable genotype.
  • T(3;7)(q27;q32) fuses BCL6 to a non-coding region at FRA7H near miR-29.

    Schneider, B; Nagel, S; Kaufmann, M; Winkelmann, S; Bode, J; Drexler, H G; MacLeod, R A F (2008-06)
  • Performance of Genomic Bordering Elements at Predefined Genomic Loci

    Goetze, Sandra; Baer, Alexandra; Winkelmann, Silke; Nehlsen, Kristina; Seibler, Jost; Maass, Karin; Bode, Jürgen (American Society for Microbiology, 2005-03)
  • Nuclear scaffold/matrix attached region modules linked to a transcription unit are sufficient for replication and maintenance of a mammalian episome

    Jenke, Andreas C. W.; Stehle, Isa M.; Herrmann, Frank; Eisenberger, Tobias; Baiker, Armin; Bode, Jürgen; Fackelmayer, Frank O.; Lipps, Hans J. (National Academy of Sciences, 2004-08-03)
  • Evaluation of sequence motifs found in scaffold/matrix-attached regions (S/MARs)

    Liebich, I.; Bode, J.; Reuter, I.; Wingender, E. (Oxford University Press, 2002-08-01)