Cyclic-di-GMP (c-di-GMP) is considered one of the most important regulators of bacterial adaptation strategies such as persistence, cytotoxicity, development and biofilm formation. C-di-GMP is synthesized by diguanylate cyclases (DGCs), containing GGDEF domain and degraded by specific phosphodiesterases (PDEs), containing EAL or HD-GYP signatures. The relevance of c-di-GMP is linked to its amazing capacity to interact with a large repertoire of proteins and nucleic acids using different binding modes; its conformational plasticity is likely at the basis of its extraordinary success as a signal molecule (1). Activaction/deactivation of individual proteins is likely to be controlled by a combination of c-di-GMP affinity and binding mode, producing a variety of allosteric control mechanisms, few of them characterized in detail biochemically. In this study, we focused on the biochemistry of c-di-GMP by studying the protein domains involved in its synthesis and degradation, which are supposed to interact to each other allosterically. To derive the principles governing protein/c-di-GMP interaction and catalysis, we have produced an array of small molecules, mostly c-di-GMP analogues, and studies their binding affinities for the active and inhibitory sites of GGDEF diguanylate cyclases (PleD, YfiN) (2,3) also in comparison with EAL phosphodiesterases (RocR) (4). Moreover, to determine the molecular determinants of the crosstalk between GGDEF and EAL domains, we focus on the characterization of the structure and function of different hybrid proteins, harboring both domains fused in tandem. 1. Romlig, U., Galperin, M.Y:, Gomelsky, M. (2013) Mol. Biol. Rev. 77:1-52. 2. Chan, C., Paul, R., Samoray, D., Amiot, N.C., Giese, B., et al (2004) Proc Natl Acad Sci USA 101: 17084-17089. 3. Giardina, G., Paiardini, A., Fernicola, S., Franceschini, S., Rinaldo, S., Stelitano, V., Cutruzzolà, F. (2013) PLoS One 8:e81324. 4. Rao, F., Yang, Y., Qi, Y., Liang, Z.X. (2008) J Bacteriol. 190:3622-31
Allosteric control in the synthesis and sensing of cyclic-di-GMP
PETRELLI, Riccardo;TORQUATI, ILARIA;CAPPELLACCI, Loredana;
2015-01-01
Abstract
Cyclic-di-GMP (c-di-GMP) is considered one of the most important regulators of bacterial adaptation strategies such as persistence, cytotoxicity, development and biofilm formation. C-di-GMP is synthesized by diguanylate cyclases (DGCs), containing GGDEF domain and degraded by specific phosphodiesterases (PDEs), containing EAL or HD-GYP signatures. The relevance of c-di-GMP is linked to its amazing capacity to interact with a large repertoire of proteins and nucleic acids using different binding modes; its conformational plasticity is likely at the basis of its extraordinary success as a signal molecule (1). Activaction/deactivation of individual proteins is likely to be controlled by a combination of c-di-GMP affinity and binding mode, producing a variety of allosteric control mechanisms, few of them characterized in detail biochemically. In this study, we focused on the biochemistry of c-di-GMP by studying the protein domains involved in its synthesis and degradation, which are supposed to interact to each other allosterically. To derive the principles governing protein/c-di-GMP interaction and catalysis, we have produced an array of small molecules, mostly c-di-GMP analogues, and studies their binding affinities for the active and inhibitory sites of GGDEF diguanylate cyclases (PleD, YfiN) (2,3) also in comparison with EAL phosphodiesterases (RocR) (4). Moreover, to determine the molecular determinants of the crosstalk between GGDEF and EAL domains, we focus on the characterization of the structure and function of different hybrid proteins, harboring both domains fused in tandem. 1. Romlig, U., Galperin, M.Y:, Gomelsky, M. (2013) Mol. Biol. Rev. 77:1-52. 2. Chan, C., Paul, R., Samoray, D., Amiot, N.C., Giese, B., et al (2004) Proc Natl Acad Sci USA 101: 17084-17089. 3. Giardina, G., Paiardini, A., Fernicola, S., Franceschini, S., Rinaldo, S., Stelitano, V., Cutruzzolà, F. (2013) PLoS One 8:e81324. 4. Rao, F., Yang, Y., Qi, Y., Liang, Z.X. (2008) J Bacteriol. 190:3622-31I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
