PilB and PilT are ATPases acting antagonistically in type IV pili function in Myxococcus xanthus

Department of Ecophysiology, Max Planck Institute for Terrestrial Microbiology, Karl-von-Frisch Str., 35043 Marburg, Germany; Georg-August-Universität Göttingen, Institute for Microbiology and Genetics, Grisebachstr. 8, 37077 Göttingen, Germany

^* To whom correspondence should be addressed. Email: sogaard{at}mpi-marburg.mpg.de.

	Abstract

Type IV pili (T4P) are dynamic surface structures that undergo cycles of extension and retraction. T4P dynamics center on the PilB and PilT proteins, which are members of the secretion ATPase superfamily of proteins. Here, we show that PilB and PilT of the T4P system in Myxococcus xanthus have ATPase activity in vitro. Using a structure-guided approach, we systematically mutagenized PilB and PilT to resolve whether both ATP binding and hydrolysis are important for PilB and PilT function in vivo. PilB as well as PilT ATPase activity was abolished in vitro by substitution of conserved residues in the Walker A and Walker B boxes involved in ATP binding and hydrolysis, respectively. PilB proteins containing mutant Walker A or Walker B boxes were non-functional in vivo and unable to support T4P extension. PilT proteins containing mutant Walker A or Walker B boxes were also non-functional in vivo and unable to support T4P retraction. These data provide genetic evidence that both ATP binding and hydrolysis by PilB are essential for T4P extension and that both ATP binding and hydrolysis by PilT are essential for T4P retraction. Thus, PilB and PilT are ATPases that act at distinct steps in the T4P extension/retraction cycle in vivo.