Divisome minimization shows that FtsZ and SepF can form an active Z-ring, and reveals BraB as a new cell division influencing protein in Bacillus subtilis

Abstract

One of the aims of synthetic biology is the construction of a minimal cell. Since bacteria are the simplest life forms, they are the preferred blueprint for such a cell. In this study, we tried to find the minimal set of proteins required for division of a cell wall containing cell, using the bacterial model system Bacillus subtilis. Bacterial cytokinesis begins with polymerization of the tubulin homologue FtsZ at midcell into the Z-ring, which recruits the late cell division protein that synthesize the division septum. Assembly of FtsZ is carefully regulated, involving a dozen conserved cell division proteins. These proteins are not essential, but removing more than one is in many cases lethal. We made use of known suppressor mutations to find a gene deletion route that eventually enabled us to remove eight conserved cell division proteins: ZapA, MinC, MinJ, UgtP, ClpX, Noc, EzrA and FtsA. Only FtsZ and its membrane anchor SepF appeared to be required for Z-ring formation. Interestingly, SepF is also the FtsZ anchor in archaea, and both proteins may date back to the Last Universal Common Ancestor (LUCA). Curiously, viability was not greatly affected by the multiple deletions, although the frequency of cell division was considerably reduced. However, genome sequencing exposed the accumulation of several suppressor mutations, and revealed an unexpected cell division regulation function for the branched chain amino acid transporter BraB. The implications of these findings for the role of SepF in cell division, and the construction of a minimal cell division machinery are discussed.