%0 Journal Article %1 perezmendoza2017novel %A Pérez-Mendoza, D. %A Bertinetti, D. %A Lorenz, R. %A Gallegos, M. %A Herberg, F. W. %A Sanjuán, J. %D 2017 %J Scientific Reports %K herberg myown %N 1 %P 8997-- %R 10.1038/s41598-017-09290-2 %T A novel c-di-GMP binding domain in glycosyltransferase BgsA is responsible for the synthesis of a mixed-linkage β-glucan %U https://doi.org/10.1038/s41598-017-09290-2 %V 7 %X BgsA is the glycosyltransferase (GT) involved in the synthesis of a linear mixed-linkage β-glucan (MLG), a recently described exopolysaccharide activated by c-di-GMP in Sinorhizobium meliloti and other Rhizobiales. Although BgsA displays sequence and structural homology with bacterial cellulose synthases (CS), it does not contain any predictable c-di-GMP binding domain. In this work we demonstrate that the cytoplasmic C-terminal domain of BgsA (C-BgsA) binds c-di-GMP with both high affinity (KD = 0.23 μM) and specificity. C-BgsA is structurally different to the otherwise equivalent cytoplasmic C-terminal domain of CS, and does not contain PilZ motifs for c-di-GMP recognition. A combination of random and site-directed mutagenesis with surface plasmon resonance (SPR) allowed identification of the C-BgsA residues which are important not only for c-di-GMP binding, but also for BgsA GT activity. The results suggest that the C-BgsA domain is important for both, c-di-GMP binding and GT activity of BgsA. In contrast to bacterial CS where c-di-GMP has been proposed as a derepressor of GT activity, we hypothesize that the C-terminal domain of BgsA plays an active role in BgsA GT activity upon binding c-di-GMP.

@article{perezmendoza2017novel, abstract = {BgsA is the glycosyltransferase (GT) involved in the synthesis of a linear mixed-linkage β-glucan (MLG), a recently described exopolysaccharide activated by c-di-GMP in Sinorhizobium meliloti and other Rhizobiales. Although BgsA displays sequence and structural homology with bacterial cellulose synthases (CS), it does not contain any predictable c-di-GMP binding domain. In this work we demonstrate that the cytoplasmic C-terminal domain of BgsA (C-BgsA) binds c-di-GMP with both high affinity (KD = 0.23 μM) and specificity. C-BgsA is structurally different to the otherwise equivalent cytoplasmic C-terminal domain of CS, and does not contain PilZ motifs for c-di-GMP recognition. A combination of random and site-directed mutagenesis with surface plasmon resonance (SPR) allowed identification of the C-BgsA residues which are important not only for c-di-GMP binding, but also for BgsA GT activity. The results suggest that the C-BgsA domain is important for both, c-di-GMP binding and GT activity of BgsA. In contrast to bacterial CS where c-di-GMP has been proposed as a derepressor of GT activity, we hypothesize that the C-terminal domain of BgsA plays an active role in BgsA GT activity upon binding c-di-GMP.}, added-at = {2017-10-13T10:39:50.000+0200}, author = {Pérez-Mendoza, D. and Bertinetti, D. and Lorenz, R. and Gallegos, M. and Herberg, F. W. and Sanjuán, J.}, biburl = {https://www.bibsonomy.org/bibtex/2081cbbe0ad8d1475e319b293e20fec3b/biochemie}, description = {A novel c-di-GMP binding domain in glycosyltransferase BgsA is responsible for the synthesis of a mixed-linkage β-glucan | Scientific Reports}, doi = {10.1038/s41598-017-09290-2}, interhash = {fd90fc609a674a350452f5afba2610f1}, intrahash = {081cbbe0ad8d1475e319b293e20fec3b}, issn = {20452322}, journal = {Scientific Reports}, keywords = {herberg myown}, number = 1, pages = {8997--}, refid = {Pérez-Mendoza2017}, timestamp = {2017-10-13T10:39:50.000+0200}, title = {A novel c-di-GMP binding domain in glycosyltransferase BgsA is responsible for the synthesis of a mixed-linkage β-glucan}, url = {https://doi.org/10.1038/s41598-017-09290-2}, volume = 7, year = 2017 }