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Journal of Bacteriology, July 2009, p. 4186-4194, Vol. 191, No. 13
0021-9193/09/$08.00+0     doi:10.1128/JB.01758-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Two-Step Assembly Dynamics of the Bacillus subtilis Divisome{triangledown} ,{dagger}

Pamela Gamba,1 Jan-Willem Veening,1 Nigel J. Saunders,2 Leendert W. Hamoen,1 and Richard A. Daniel1*

Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University, Medical School, Framlington Place, Newcastle-upon-Tyne NE2 4HH, United Kingdom,1 Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, United Kingdom2

Received 16 December 2008/ Accepted 24 April 2009

Cell division in bacteria is carried out by about a dozen proteins which assemble at midcell and form a complex known as the divisome. To study the dynamics and temporal hierarchy of divisome assembly in Bacillus subtilis, we have examined the in vivo localization pattern of a set of division proteins fused to green fluorescent protein in germinating spores and vegetative cells. Using time series and time-lapse microscopy, we show that the FtsZ ring assembles early and concomitantly with FtsA, ZapA, and EzrA. After a time delay of at least 20% of the cell cycle, a second set of division proteins, including GpsB, FtsL, DivIB, FtsW, Pbp2B, and DivIVA, are recruited to midcell. Together, our data provide in vivo evidence for two-step assembly of the divisome. Interestingly, overproduction of FtsZ advances the temporal assembly of EzrA but not of DivIVA, suggesting that a signal different from that of FtsZ polymerization drives the assembly of late divisome proteins. Microarray analysis shows that FtsZ depletion or overexpression does not significantly alter the transcription of division genes, supporting the hypothesis that cell division in B. subtilis is mainly regulated at the posttranscriptional level.

* Corresponding author. Mailing address: Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University, Medical School, Framlington Place, Newcastle-upon-Tyne NE2 4HH, United Kingdom. Phone: 0191 2228866. Fax: 0191 2227424. E-mail: richard.daniel{at}ncl.ac.uk

{triangledown} Published ahead of print on 8 May 2009.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, July 2009, p. 4186-4194, Vol. 191, No. 13
0021-9193/09/$08.00+0     doi:10.1128/JB.01758-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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