White Rose University Consortium logo
University of Leeds logo University of Sheffield logo York University logo

Dimer-induced signal propagation in Spo0A

Muchova, K, Lewis, R J, Perecko, D, Brannigan, J A, Ladds, J C, Leech, A, Wilkinson, A J (orcid.org/0000-0003-4577-9479) and Barak, I (2004) Dimer-induced signal propagation in Spo0A. Molecular Microbiology. pp. 829-842. ISSN 0950-382X

Text (branniganja6.pdf)

Download (375Kb)


Spo0A, the response regulator protein controlling the initiation of sporulation in Bacillus, has two distinct domains, an N-terminal phosphoacceptor (or receiver) domain and a C-terminal DNA-binding (or effector) domain. The phosphoacceptor domain mediates dimerization of Spo0A on phosphorylation. A comparison of the crystal structures of phosphorylated and unphosphorylated response regulators suggests a mechanism of activation in which structural changes originating at the phosphorylatable aspartate extend to the alpha(4)beta(5)alpha(5) surface of the protein. In particular, the data show an important role in downstream signalling for a conserved aromatic residue (Phe-105 in Spo0A), the conformation of which alters upon phosphorylation. In this study, we have prepared a Phe-105 to Ala mutant to probe the contribution of this residue to Spo0A function. We have also made an alanine substitution of the neighbouring residue Tyr-104 that is absolutely conserved in the Spo0As of spore-forming Bacilli. The spo0A(Y104A) and spo0A(F105A) alleles severely impair sporulation in vivo. In vitro phosphorylation of the purified proteins by phosphoramidate is unaffected, but dimerization and DNA binding are abolished by the mutations. We have identified intragenic suppressor mutations of spo0A(F105A) and shown that these second-site mutations in the purified proteins restore phosphorylation-dependent dimer formation. Our data support a model in which dimerization and signal transduction between the two domains of Spo0A are mediated principally by the alpha(4)beta(5)alpha(5) signalling surface in the receiver domain.

Item Type: Article
Copyright, Publisher and Additional Information: © 2004 Blackwell Publishing Ltd. This is an electronic version of an article published in Molecular Microbiology: complete citation information for the final version of the paper, as published in the print edition of Molecular Microbiology, is available on the Blackwell Synergy online delivery service, accessible via the journal's website at http://www.blackwellpublishing.com/journal.asp?ref=0950-382X or www.blackwell-synergy.com
Institution: The University of York
Academic Units: The University of York > Chemistry (York)
Depositing User: Repository Officer
Date Deposited: 29 Apr 2005
Last Modified: 21 Aug 2016 00:05
Published Version: http://dx.doi.org/10.1111/j.1365-2958.2004.04171.x
Status: Published
Refereed: Yes
URI: http://eprints.whiterose.ac.uk/id/eprint/448

Actions (repository staff only: login required)