Universität zu Köln

Perez-Mendoza, Daniel, Bertinetti, Daniela ORCID: 0000-0003-1150-3765, Lorenz, Robin, Gallegos, Maria-Trinidad, Herberg, Friedrich W. and Sanjuan, Juan (2017). A novel c-di-GMP binding domain in glycosyltransferase BgsA is responsible for the synthesis of a mixed-linkage beta-glucan. Sci Rep, 7. LONDON: NATURE PUBLISHING GROUP. ISSN 2045-2322

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Abstract

BgsA is the glycosyltransferase (GT) involved in the synthesis of a linear mixed-linkage beta-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 (K-D = 0.23 mu M) 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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Perez-Mendoza, Daniel UNSPECIFIED UNSPECIFIED UNSPECIFIED Bertinetti, Daniela UNSPECIFIED orcid.org/0000-0003-1150-3765 UNSPECIFIED Lorenz, Robin UNSPECIFIED UNSPECIFIED UNSPECIFIED Gallegos, Maria-Trinidad UNSPECIFIED UNSPECIFIED UNSPECIFIED Herberg, Friedrich W. UNSPECIFIED UNSPECIFIED UNSPECIFIED Sanjuan, Juan UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-221590
DOI:	10.1038/s41598-017-09290-2
Journal or Publication Title:	Sci Rep
Volume:	7
Date:	2017
Publisher:	NATURE PUBLISHING GROUP
Place of Publication:	LONDON
ISSN:	2045-2322
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language BACTERIAL CELLULOSE SYNTHASE; PILZ-DOMAIN; PSEUDOMONAS-AERUGINOSA; SIGNAL-TRANSDUCTION; ALLOSTERIC CONTROL; BIOFILM FORMATION; STRUCTURAL BASIS; RECEPTOR; REGULATOR; MECHANISM Multiple languages Multidisciplinary Sciences Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/22159