Evidence for a second regulatory binding site on PspF that is occupied by the C-terminal domain of PspA

authored by: Eyleen Sabine Heidrich, Thomas Brüser
Abstract: PspA is a key component of the bacterial Psp membrane-stress response system. The biochemical and functional characterization of PspA is impeded by its oligomerization and aggregation properties. It was recently possible to solve the coiled coil structure of a completely soluble PspA fragment, PspA(1–144), that associates with the σ⁵⁴ enhancer binding protein PspF at its W56-loop and thereby down-regulates the Psp response. We now found that the C-terminal part of PspA, PspA(145–222), also interacts with PspF and inhibits its activity in the absence of full-length PspA. Surprisingly, PspA(145–222) effects changed completely in the presence of full-length PspA, as promoter activity was triggered instead of being inhibited under this condition. PspA(145–222) thus interfered with the inhibitory effect of full-length PspA on PspF, most likely by interacting with full-length PspA that remained bound to PspF. In support of this view, a comprehensive bacterial-2-hybrid screen as well as co-purification analyses indicated a self-interaction of PspA(145–222) and an interaction with full-length PspA. This is the first direct demonstration of PspA/PspA and PspA/PspF interactions in vivo that are mediated by the C-terminus of PspA. The data indicate that regulatory binding sites on PspF do not only exist for the N-terminal coiled coil domain but also for the C-terminal domain of PspA. The inhibition of PspF by PspA-(145–222) was reduced upon membrane stress, whereas the inhibition of PspF by PspA(1–144) did not respond to membrane stress. We therefore propose that the C-terminal domain of PspA is crucial for the regulation of PspF in response to Psp system stimuli.
Organisation(s): Institute of Microbiology
Type: Article
Journal: PLOS ONE
Volume: 13
ISSN: 1932-6203
Publication date: 15.06.2018
Publication status: Published
Peer reviewed: Yes
ASJC Scopus subject areas: General Biochemistry,Genetics and Molecular Biology, General Agricultural and Biological Sciences, General
Electronic version(s): https://doi.org/10.1371/journal.pone.0198564 (Access: Open)
https://doi.org/10.15488/4715 (Access: Open)