Open AccessAnalysis of the HD-GYP Domain Cyclic Dimeric GMP Phosphodiesterase Reveals a Role in Motility and the Enzootic Life Cycle of Borrelia burgdorferi
Author(s) -
Syed Z. Sultan,
Joshua E. Pitzer,
Tristan Boquoi,
Gerry Hobbs,
Michael R. Miller,
M. A. Motaleb
Publication year - 2011
Publication title -
infection and immunity
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.508
H-Index - 220
eISSN - 1070-6313
pISSN - 0019-9567
DOI - 10.1128/iai.05153-11
Subject(s) - biology , phosphodiesterase , borrelia burgdorferi , motility , mutant , virulence , microbiology and biotechnology , chemotaxis , biochemistry , enzyme , receptor , genetics , gene , antibody
HD-GYP domain cyclic dimeric GMP (c-di-GMP) phosphodiesterases are implicated in motility and virulence in bacteria.Borrelia burgdorferi possesses a single set of c-di-GMP-metabolizing enzymes, including a putative HD-GYP domain protein, BB0374. Recently, we characterized the EAL domain phosphodiesterase PdeA. A mutation inpdeA resulted in cells that were defective in motility and virulence. Here we demonstrate that BB0374/PdeB specifically hydrolyzed c-di-GMP with aKm of 2.9 nM, confirming that it is a functional phosphodiesterase. Furthermore, by measuring phosphodiesterase enzyme activity in extracts from cells containing thepdeA pdeB double mutant, we demonstrate that no additional phosphodiesterases are present inB. burgdorferi .pdeB single mutant cells exhibit significantly increased flexing, indicating a role for c-di-GMP in motility. Constructing and analyzing apilZ pdeB double mutant suggests that PilZ likely interacts with chemotaxis signaling. While virulence in needle-inoculated C3H/HeN mice did not appear to be altered significantly inpdeB mutant cells, these cells exhibited a reduced ability to survive inIxodes scapularis ticks. Consequently, those ticks were unable to transmit the infection to naïve mice. All of these phenotypes were restored when the mutant was complemented. Identification of this role ofpdeB increases our understanding of the c-di-GMP signaling network in motility regulation and the life cycle ofB. burgdorferi .