Systematic Identification of Cyclic-di-GMP Binding Proteins in Vibrio cholerae Reveals a Novel Class of Cyclic-di-GMP-Binding ATPases Associated with Type II Secretion Systems

dc.contributor.authorRoelofs, Kevin G.
dc.contributor.authorJones, Christopher J.
dc.contributor.authorHelman, Sarah R.
dc.contributor.authorShang, Xiaoran
dc.contributor.authorOrr, Mona W.
dc.contributor.authorGoodson, Jonathan R.
dc.contributor.authorGalperin, Michael Y.
dc.contributor.authorYildiz, Fitnat H.
dc.contributor.authorLee, Vincent T.
dc.date.accessioned2017-08-30T18:36:42Z
dc.date.available2017-08-30T18:36:42Z
dc.date.issued2015-10-27
dc.descriptionFunding for Open Access provided by the UMD Libraries Open Access Publishing Fund.en_US
dc.description.abstractCyclic-di-GMP (c-di-GMP) is a ubiquitous bacterial signaling molecule that regulates a variety of complex processes through a diverse set of c-di-GMP receptor proteins. We have utilized a systematic approach to identify c-di-GMP receptors from the pathogen Vibrio cholerae using the Differential Radial Capillary Action of Ligand Assay (DRaCALA). The DRaCALA screen identified a majority of known c-di-GMP binding proteins in V. cholera and revealed a novel c-di-GMP binding protein, MshE (VC0405), an ATPase associated with the mannose sensitive hemagglutinin (MSHA) type IV pilus. The known c-di-GMP binding proteins identified by DRaCALA include diguanylate cyclases, phosphodiesterases, PilZ domain proteins and transcription factors VpsT and VpsR, indicating that the DRa- CALA-based screen of open reading frame libraries is a feasible approach to uncover novel receptors of small molecule ligands. Since MshE lacks the canonical c-di-GMP-binding motifs, a truncation analysis was utilized to locate the c-di-GMP binding activity to the N-terminal T2SSE_N domain. Alignment of MshE homologs revealed candidate conserved residues responsible for c-di-GMP binding. Site-directed mutagenesis of these candidate residues revealed that the Arg9 residue is required for c-di-GMP binding. The ability of c-di-GMP binding to MshE to regulate MSHA dependent processes was evaluated. The R9A allele, in contrast to the wild type MshE, was unable to complement the ΔmshE mutant for the production of extracellular MshA to the cell surface, reduction in flagella swimming motility, attachment to surfaces and formation of biofilms. Testing homologs of MshE for binding to c-di-GMP identified the type II secretion ATPase of Pseudomonas aeruginosa (PA14_29490) as a c-di-GMP receptor, indicating that type II secretion and type IV pili are both regulated by c-di-GMP.en_US
dc.identifierhttps://doi.org/10.13016/M2SX64940
dc.identifier.citationRoelofs KG, Jones CJ, Helman SR, Shang X, Orr MW, Goodson JR, et al. (2015) Systematic Identification of Cyclic-di-GMP Binding Proteins in Vibrio cholerae Reveals a Novel Class of Cyclic-di- GMP-Binding ATPases Associated with Type II Secretion Systems. PLoS Pathog 11(10): e1005232. doi:10.1371/journal.ppat.1005232en_US
dc.identifier.urihttp://hdl.handle.net/1903/19684
dc.language.isoen_USen_US
dc.publisherPLOS (Public Library of Science)en_US
dc.relation.isAvailableAtCell Biology & Molecular Geneticsen_us
dc.relation.isAvailableAtDigital Repository at the University of Marylanden_us
dc.relation.isAvailableAtCollege of Computer, Mathematical & Natural Sciencesen_us
dc.relation.isAvailableAtUniversity of Maryland (College Park, MD)en_us
dc.titleSystematic Identification of Cyclic-di-GMP Binding Proteins in Vibrio cholerae Reveals a Novel Class of Cyclic-di-GMP-Binding ATPases Associated with Type II Secretion Systemsen_US
dc.typeArticleen_US

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