GIL, a new c‐di‐GMP‐binding protein domain involved in regulation of cellulose synthesis in enterobacteria. Issue 3 (1st July 2014)

• Record Type: Journal Article
• Authors: Fang, Xin; Ahmad, Irfan; Blanka, Andrea; Schottkowski, Marco; Cimdins, Annika; Galperin, Michael Y.; Römling, Ute; Gomelsky, Mark
• Is Part Of: Molecular microbiology. Volume 93:Issue 3(2014)
• Page Start: 439
• Page End: 452
• Publication Date: 2014-07-01
• Subjects: Molecular microbiology -- Periodicals; 572.829
• DOI: 10.1111/mmi.12672
• Languages: English
• ISSNs: 0950-382X

Abstract

<abstract abstract-type="main"> <title>Summary</title> <p>In contrast to numerous enzymes involved in c‐di‐GMP synthesis and degradation in enterobacteria, only a handful of c‐di‐GMP receptors/effectors have been identified. In search of new c‐di‐GMP receptors, we screened the <italic>E</italic><italic>scherichia coli</italic> ASKA overexpression gene library using the Differential Radial Capillary Action of Ligand Assay (DRaCALA) with fluorescently and radioisotope‐labelled c‐di‐GMP. We uncovered three new candidate c‐di‐GMP receptors in <italic>E</italic><italic>. coli</italic> and characterized one of them, BcsE. The <italic>bcsE</italic> gene is encoded in cellulose synthase operons in representatives of Gammaproteobacteria and Betaproteobacteria. The purified BcsE proteins from <italic>E</italic><italic>. coli</italic>, <italic>S</italic><italic>almonella enterica</italic> and <italic>K</italic><italic>lebsiella pneumoniae</italic> bind c‐di‐GMP via the domain of unknown function, DUF2819, which is hereby designated GIL, <underline>G</underline>GDEF <underline>I</underline>‐site <underline>l</underline>ike domain. The RxGD motif of the GIL domain is required for c‐di‐GMP binding, similar to the c‐di‐GMP‐binding I‐site of the diguanylate cyclase GGDEF domain. Thus, GIL is the second protein domain, after PilZ, dedicated to c‐di‐GMP‐binding. We show that in <italic>S.</italic> <italic>enterica</italic>, BcsE is not essential for cellulose synthesis but is required for maximal cellulose production, and that c‐di‐GMP binding is critical for BcsE function. It appears that cellulose production in enterobacteria is controlled by a two‐tiered c‐di‐GMP‐dependent system involving BcsE and the PilZ domain containing glycosyltransferase BcsA.</p> </abstract>