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Low-copy-number plasmids require sophisticated genetic devices to achieve efficient segregation of plasmid copies during cell division. Plasmid R388 uses a unique segregation mechanism, based on StbA, a small multifunctional protein. StbA is the key protein in a segregation system not involving a plasmid-encoded NTPase partner, it regulates the expression of several plasmid operons, and it is the main regulator of plasmid conjugation. The mechanisms by which StbA, together with the centromere-like sequence stbS, achieves segregation, is largely uncharacterized. To better understand the molecular basis of R388 segregation, we determined the crystal structure of the conserved N-terminal domain of StbA to 1.9 Å resolution. It folds into an HTH DNA-binding domain, structurally related to that of the PadR subfamily II of transcriptional regulators. StbA is organized in two domains. Its N-terminal domain carries the specific stbS DNA binding activity. A truncated version of StbA, deleted of its C-terminal domain, displays only partial activities in vivo, indicating that the non-conserved C-terminal domain is required for efficient segregation and subcellular plasmid positioning. The structure of StbA DNA-binding domain also provides some insight into how StbA monomers cooperate to repress transcription by binding to the stbDR and to form the segregation complex with stbS., This work was supported by the Spanish Ministry of Economy, Industry and Competitiveness grant BFU2017-86378-P to F.dlC., by the Spanish Ministry of Science (MCI/AEI/FEDER,UE) grant PGC2018-093885-B-I00 to G.M., by French National Research Agency, grant numberANR-18- CE35-0008 to J.-Y.B. and by University P. Sabatier grant to C.G.

In this study, we analyzed and developed a phosphor with a nitridiosilicate based and emitting color changing from green to orange to apply to our white-light-emitting diodes (WLEDs). Eu2+ and Yb2+ trap sites for twodoped SrSi2O2N2:Eu2+,Yb2+ (SrYb) nitridosilicate emit a wide emission in the region between green and orange. By calculating the decompose time of the green-emission power donor, we could determine the converted energy between the active ions. Furthermore, we also analyzed the impact of codoping with varied active ion ratios on the photoluminescence (PL) characteristics. As the concentration of Yb2+ increases, the red emission dominates the green. This was because the obtained phoshor's emission depended on the activator ion components. To generate white light, a combination containing the acquired phosphor and the indium gallium nitride (InGaN) blue LED chip with 450-nm wavelength was used. With only two steps, we could detect the hue balance management. Firstly, by modifying the Eu2+ and Yb2+ concentrations, we can conduct a green-toorange proportion optimization. Following that, the commission on illumination (CIE) coordinates were transformed to [0.4071; 0.3789] from the original position, which was [0.2805; 0.2014] by enhancing the phosphor powder amount. An environment which has a CRI of around 89 is the optimum condition for conducting white light.

REP, diverse palindromic DNA sequences found at high copy number in many bacterial genomes, have been attributed important roles in cell physiology but their dissemination mechanisms are poorly understood. They might represent non-autonomous transposable elements mobilizable by TnpAREP, the first prokaryotic domesticated transposase associated with REP. TnpAREP, fundamentally different from classical transposases, are members of the HuH superfamily and closely related to the transposases of the IS200/IS605 family. We previously showed that Escherichia coli TnpAREP processes cognate single stranded REP in vitro and that this activity requires the integrity of the REP structure, in particular imperfect palindromes interrupted by a bulge and preceded by a conserved DNA motif. A second group of REPs rather carry perfect palindromes, raising questions about how the latter are recognized by their cognate TnpAREP. To get insight into the importance of REP structural and sequence determinants in these two groups, we developed an in vitro activity assay coupled to a mutational analysis for three different TnpAREP/REP duos via a SELEX approach. We also tackled the question of how the cleavage site is selected. This study revealed that two TnpAREP groups have co-evolved with their cognate REPs and use different strategies to recognize their REP substrates.

Bacterial transposons, through their ability to transfer DNA sequences from one position in the genome to another, play a central role in the shape and the evolution of genomes. Extensive studies have been performed during the last five decades to understand the molecular mechanisms involved in the transposition of a variety of elements. Among the methods used, the papillation and the mating out coupled to arbitrary primed PCR assays described in this chapter are widely used as very powerful approaches to detect and characterize transposition events in vivo.