Structure of the PilZ-FimXEAL-c-di-GMP complex responsible for the regulation of bacterial Type IV pilus biogenesis

GUZZO, CRISTIANE; DUNGER, GERMÁN; KOPKE SALINAS, ROBERTO; FARAH, C.S.

JOURNAL OF MOLECULAR BIOLOGY

ACADEMIC PRESS LTD-ELSEVIER SCIENCE LTD

Lugar: Amsterdam; Año: 2013 vol. 425 p. 2174 - 2197

0022-2836

Signal transduction pathways mediated by cyclic-bis(3´-5´)-dimeric GMP (c-di-GMP) control many important and complex behaviors in bacteria. C-di-GMP is synthesized through the action of GGDEF domains that possess diguanylate cyclase activity and is degraded by EAL or HD-GYP domains with phosphodiesterase activity. There is mounting evidence that some important c-di-GMP-mediated pathways require protein-protein interactions between members of the GGDEF, EAL, HD-GYP and PilZ protein domain families. For example, interactions have been observed between PilZ and the EAL domain from FimX of Xanthomonas citri (Xac). FimX and PilZ are involved in the regulation of type IV pilus biogenesis via interactions of the latter with the hexameric PilB ATPase associated with the bacterial inner membrane. Here, we present the crystal structure of the ternary complex made up of PilZ, the FimX EAL domain (FimX-EAL) and c-di-GMP. PilZ interacts principally with the lobe region and the N-terminal linker helix of the FimX-EAL. These interactions involve a hydrophobic surface made up of amino acids conserved in a non-canonical family of PilZ domains that lack intrinsic c-di-GMP binding ability and strand complementation that joins b-sheets from both proteins. Interestingly, the c-di-GMP binds to isolated FimX-EAL and to the PilZ-FimX-EAL complex in a novel conformation encountered in c-di-GMP protein complexes in which one of the two glycosidic bonds is in a rare syn conformation while the other adopts the more common anti conformation. The structure points to a means by which c-di-GMP and PilZ binding could be coupled to FimX and PilB conformational states.