Poster Presentation Australian Society for Microbiology Annual Scientific Meeting 2014

IDENTIFYING HOW BACTERIA FIND THEIR MIDDLE: REOPENING THE QUESTION (#254)

Isabella Hajduk 1 , Hajime Okumura 1 , Elizabeth Harry 1
  1. The ithree institute, University of Technology Sydney (UTS), Sydney, NSW, Australia

In bacteria, the earliest stage of cell division is the formation of the Z ring at the division site at the cell centre.  It has been shown in Bacillus subtilis that Z ring assembly is coupled with the early stages of DNA replication1, 2. The chromosome (nucleoid) also influences the position of the division site via the actions of Noc (nucleoid occlusion) which binds the chromosome and prevents Z ring assembly over it too early in the replication round3. Both systems ensure each daughter cell receives a complete genetic complement. When DNA replication is blocked at the earliest stage of DNA replication (initiation), midcell Z rings are unable to form and this is partly due to the activity of Noc4. However, another DNA-binding protein, Spo0J, has been shown to influence Z ring placement5. Therefore, we hypothesised that Spo0J may have a nucleoid occlusion function. To test this, mutant strains lacking spo0J were constructed. Initiation of DNA replication was inhibited during the first round of replication using the spore outgrowth system and Z rings were observed in live cells using an ftsZ-yfp fusion and fluorescence microscopy. A significant increase in the frequency of midcell Z rings was observed in cells lacking Spo0J when initiation was blocked, to a similar frequency previously observed in cells lacking Noc. These results support our hypothesis of Spo0J having a nucleoid occlusion function. We are now testing whether removing both Noc and Spo0J would result in complete midcell Z ring assembly when initiation of DNA replication is blocked. If so, this would connect the nucleoid occlusion proteins with the coupling of DNA replication and division site placement.

  1. Harry, E.J., Rodwell, J., and Wake, R.G. (1999). Co-ordinating DNA replication with cell division in bacteria: a link between the early stages of a round of replication and mid-cell Z ring assembly. Molecular Microbiology 33, 33-40.
  2. Regamey, A., Harry, E.J., and Wake, R.G. (2000). Mid-cell Z ring assembly in the absence of entry into the elongation phase of the round of replication in bacteria: co-ordinating chromosome replication with cell division. Molecular Microbiology 38, 423-434.
  3. Wu, L.J., and Errington, J. (2004). Coordination of Cell Division and Chromosome Segregation by a Nucleoid Occlusion Protein in Bacillus subtilis. Cell 117, 915-925.
  4. Moriya, S., Rashid, R.A., Rodrigues, C.D.A., and Harry, E.J. (2010). Influence of the nucleoid and the early stages of DNA replication on positioning the division site in Bacillus subtilis. Molecular Microbiology 76, 634-647.
  5. Ireton, K., Gunther, N.W., 4th, and Grossman, A.D. (1994). spo0J is required for normal chromosome segregation as well as the initiation of sporulation in Bacillus subtilis. Journal of Bacteriology 176, 5320-5329