Abstract
Diguanylate cyclases that synthesize and phosphodiesterases that hydrolyze the second messenger cyclic-di-GMP (c-di-GMP) are at the center of bacterial signaling pathways that control behaviors relevant to all aspects of microbial physiology and pathogenesis (Romling et al., Microbiol Mol Biol Rev 77(1):1–52, 2013). Bioinformatics tools can easily predict the presence of the diguanylate cyclase GGDEF domain, or the EAL and HD-GYP domains associated with phosphodiesterase activity. However, experimental confirmation of enzymatic activity is still necessary, as many proteins contain degenerate domains that lack catalytic activity but nonetheless function as c-di-GMP receptors.
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References
Romling U, Galperin MY, Gomelsky M (2013) Cyclic di-GMP: the first 25 years of a universal bacterial second messenger. Microbiol Mol Biol Rev 77(1):1–52. doi:10.1128/mmbr.00043-12
Aldridge P, Paul R, Goymer P, Rainey PB, Jenal U (2003) Role of the GGDEF regulator PleD in polar development of Caulobacter crescentus. Mol Microbiol 47:1695–1708, DOI: 10.1046/j.1365-2958.2003.03401.x
Paul R, Weiser S, Amiot NC, Chan C, Schirmer T, Giese B, Jenal U (2004) Cell cycle-dependent dynamic localization of a bacterial response regulator with a novel di-guanylate cyclase output domain. Genes Dev 18:715–727. doi:10.1101/gad.289504
Jain R, Behrens AJ, Kaever V, Kazmiercak BI (2012) Type IV pilus assembly in Pseudomonas aeruginosa over a broad range of cyclic-di-GMP concentrations. J Bacteriol 194:4285–4294. doi:10.1128/JB.00803-12
Bornhorst J, Falke J (2000) Purification of proteins using polyhistidine affinity tags. Methods Enzymol 326:245–254
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Kazmierczak, B.I. (2017). Synthesis of [32P]-c-di-GMP for Diguanylate Cyclase and Phosphodiesterase Activity Determinations. In: Sauer, K. (eds) c-di-GMP Signaling. Methods in Molecular Biology, vol 1657. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7240-1_3
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DOI: https://doi.org/10.1007/978-1-4939-7240-1_3
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-7240-1
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